Antagonists of chemokine receptors

ABSTRACT

Compounds are provided that act as potent antagonists of the CCR1 receptor, and have in vivo anti-inflammatory activity. The compounds are generally aryl piperazine derivatives and are useful in pharmaceutical compositions, methods for the treatment of CCR1-mediated diseases, and as controls in assays for the identification of competitive CCR1 antagonists.

CROSS-REFERENCES TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.14/878,818 filed Oct. 8, 2015, which is a divisional of U.S. applicationSer. No. 14/011,174 filed Aug. 27, 2013 (now U.S. Pat. No. 9,181,241),which claims the benefit of priority to U.S. Application Ser. No.61/693,758, filed Aug. 27, 2012 and 61/831,694, filed Jun. 6, 2013, thedisclosures of which are incorporated herein by reference in theirentirety.

STATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSOREDRESEARCH OR DEVELOPMENT

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REFERENCE TO A “SEQUENCE LISTING,” A TABLE, OR A COMPUTER PROGRAMLISTING APPENDIX SUBMITTED ON A COMPACT DISK

NOT APPLICABLE

BACKGROUND OF THE INVENTION

The present invention provides compounds, pharmaceutical compositionscontaining one or more of those compounds or their pharmaceuticallyacceptable salts, which are effective in inhibiting the binding ofvarious chemokines, such as MIP-1α, leukotactin, MPIF-1 and RANTES, tothe CCR1 receptor. As antagonists or modulators for the CCR1 receptor,the compounds and compositions have utility in treating inflammatory andimmune disorder conditions and diseases.

Human health depends on the body's ability to detect and destroy foreignpathogens that might otherwise take valuable resources from theindividual and/or induce illness. The immune system, which comprisesleukocytes (white blood cells (WBCs): T and B lymphocytes, monocytes,macrophages granulocytes, NK cell, mast cells, dendritic cell, andimmune derived cells (for example, osteoclasts)), lymphoid tissues andlymphoid vessels, is the body's defense system. To combat infection,white blood cells circulate throughout the body to detect pathogens.Once a pathogen is detected, innate immune cells and cytotoxic T cellsin particular are recruited to the infection site to destroy thepathogen. Chemokines act as molecular beacons for the recruitment andactivation of immune cells, such as lymphocytes, monocytes andgranulocytes, identifying sites where pathogens exist.

Despite the immune system's regulation of pathogens, certaininappropriate chemokine signaling can develop and has been attributed totriggering or sustaining inflammatory disorders, such as rheumatoidarthritis, multiple sclerosis and others. For example, in rheumatoidarthritis, unregulated chemokine accumulation in bone joints attractsand activates infiltrating macrophages and T-cells. The activities ofthese cells induce synovial cell proliferation that leads, at least inpart, to inflammation and eventual bone and cartilage loss (see,DeVries, M. E., et al., Semin Immunol 11(2):95-104 (1999)). A hallmarkof some demyelinating diseases such as multiple sclerosis is thechemokine-mediated monocyte/macrophage and T cell recruitment to thecentral nervous system (see, Kennedy, et al., J. Clin. Immunol.19(5):273-279 (1999)). Chemokine recruitment of destructive WBCs totransplants has been implicated in their subsequent rejection. See,DeVries, M. E., et al., ibid. Because chemokines play pivotal roles ininflammation and lymphocyte development, the ability to specificallymanipulate their activity has enormous impact on ameliorating andhalting diseases that currently have no satisfactory treatment. Inaddition, transplant rejection may be minimized without the generalizedand complicating effects of costly immunosuppressive pharmaceuticals.

Chemokines, a group of greater than 40 small peptides (7-10 kD), ligatereceptors expressed primarily on WBCs or immune derived cells, andsignal through G-protein-coupled signaling cascades to mediate theirchemoattractant and chemostimulant functions. Receptors may bind morethan one ligand; for example, the receptor CCR1 ligates RANTES(regulated on activation normal T cell expressed), MIP-1α (macrophageinflammatory protein), MPIF-1/CKβ8, and Leukotactin chemokines (amongothers with lesser affinities). To date, 24 chemokine receptors areknown. The sheer number of chemokines, multiple ligand bindingreceptors, and different receptor profiles on immune cells allow fortightly controlled and specific immune responses. See, Rossi, et al.,Ann. Rev. Immunol. 18(1):217-242 (2000). Chemokine activity can becontrolled through the modulation of their corresponding receptors,treating related inflammatory and immunological diseases and enablingorgan and tissue transplants.

The receptor CCR1 and its chemokine ligands, including, for exampleMIP-1α, MPIF-1/CKβ8, leukotactin and RANTES, represent significanttherapeutic targets (see Saeki, et al., Current Pharmaceutical Design9:1201-1208 (2003)) since they have been implicated in rheumatoidarthritis, transplant rejection (see, DeVries, M. E., et al., ibid.),and multiple sclerosis (see, Fischer, et al., J Neuroimmunol.110(1-2):195-208 (2000); Izikson, et al., J. Exp. Med. 192(7): 1075-1080(2000); and Rottman, et al., Eur. J. Immunol. 30(8):2372-2377 (2000). Infact, function-blocking antibodies, modified chemokine receptor ligandsand small organic compounds have been discovered, some of which havebeen successfully demonstrated to prevent or treat somechemokine-mediated diseases (reviewed in Rossi, et al., ibid.). Notably,in an experimental model of rheumatoid arthritis, disease development isdiminished when a signaling-blocking, modified-RANTES ligand isadministered (see Plater-Zyberk, et al., Immunol Lett. 57(1-3): 117-120(1997)). While function-blocking antibody and small peptide therapiesare promising, they suffer from the perils of degradation, extremelyshort half-lives once administered, and prohibitive expense to developand manufacture, characteristic of most proteins. Small organiccompounds are preferable since they often have longer half lives invivo, require fewer doses to be effective, can often be administeredorally, and are consequently less expensive. Some organic antagonists ofCCR1 have been previously described (see, Hesselgesser, et al., J. Biol.Chem. 273(25): 15687-15692 (1998); Ng, et al., J. Med. Chem.42(22):4680-4694 (1999); Liang, et al., J. Biol. Chem.275(25):19000-19008 (2000); and Liang, et al., Eur. J. Pharmacol.389(1):41-49 (2000)). In view of the effectiveness demonstrated fortreatment of disease in animal models (see, Liang, et al., J. Biol.Chem. 275(25): 19000-19008 (2000)), the search has continued to identifyadditional compounds that can be used in the treatment of diseasesmediated by CCR1 signaling.

BRIEF SUMMARY OF THE INVENTION

Described herein are compounds having the formula:

and salts, rotamers and optical isomers thereof, wherein the subscriptn, and the substituents R^(1a), R^(1b), R^(2a), R^(2b), Ar¹ and Ar² havethe meanings provided in the description and claims.

Selected groups of compounds are those of formulae Ia, Ia1, Ia2, II, IIIand IV:

In addition to the compounds provided herein, the present inventionfurther provides pharmaceutical compositions containing one or more ofthese compounds, as well as methods for the use of these compounds intherapeutic methods, primarily to treat diseases associated with CCR1signaling activity.

BRIEF DESCRIPTION OF THE DRAWINGS

NONE

DETAILED DESCRIPTION OF THE INVENTION I. Abbreviation and Definitions

The term “alkyl”, by itself or as part of another substituent, means,unless otherwise stated, a straight or branched chain hydrocarbonradical, having the number of carbon atoms designated (i.e. C₁₋₈ meansone to eight carbons). Examples of alkyl groups include methyl, ethyl,n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, n-pentyl,n-hexyl, n-heptyl, n-octyl, and the like. The term “alkenyl” refers toan unsaturated alkyl group having one or more double bonds. Similarly,the term “alkynyl” refers to an unsaturated alkyl group having one ormore triple bonds. Examples of such unsaturated alkyl groups includevinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl),2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl,3-butynyl, and the higher homologs and isomers. The term “cycloalkyl”refers to hydrocarbon rings having the indicated number of ring atoms(e.g., C₃₋₆cycloalkyl) and being fully saturated or having no more thanone double bond between ring vertices. “Cycloalkyl” is also meant torefer to bicyclic and polycyclic hydrocarbon rings such as, for example,bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane, etc. The term“heterocycloalkyl” refers to a cycloalkyl group that contain from one tofive heteroatoms selected from N, O, and S, wherein the nitrogen andsulfur atoms are optionally oxidized, and the nitrogen atom(s) areoptionally quaternized. The heterocycloalkyl may be a monocyclic, abicyclic or a polycylic ring system. Non limiting examples ofheterocycloalkyl groups include pyrrolidine, piperidinyl, imidazolidine,pyrazolidine, butyrolactam, valerolactam, imidazolidinone, hydantoin,dioxolane, phthalimide, piperidine, 1,4-dioxane, morpholine,thiomorpholine, thiomorpholine-S-oxide, thiomorpholine-S,S-oxide,piperazine, pyran, pyridone, 3-pyrroline, thiopyran, pyrone,tetrahydrofuran, tetrahydrothiophene, quinuclidine, and the like. Aheterocycloalkyl group can be attached to the remainder of the moleculethrough a ring carbon or a heteroatom.

The term “alkylene” by itself or as part of another substituent means adivalent radical derived from an alkane, as exemplified by—CH₂CH₂CH₂CH₂—. Typically, an alkyl (or alkylene) group will have from 1to 24 carbon atoms, with those groups having 10 or fewer carbon atomsbeing preferred in the present invention. A “lower alkyl” or “loweralkylene” is a shorter chain alkyl or alkylene group, generally havingfour or fewer carbon atoms. Similarly, “alkenylene” and “alkynylene”refer to the unsaturated forms of “alkylene” having double or triplebonds, respectively.

The terms “alkoxy,” “alkylamino” and “alkylthio” (or thioalkoxy) areused in their conventional sense, and refer to those alkyl groupsattached to the remainder of the molecule via an oxygen atom, an aminogroup, or a sulfur atom, respectively. Additionally, for dialkylaminogroups, the alkyl portions can be the same or different and can also becombined to form a 3-7 membered ring with the nitrogen atom to whicheach is attached. Accordingly, a group represented as —NR^(a)R^(b) ismeant to include piperidinyl, pyrrolidinyl, morpholinyl, azetidinyl andthe like.

The terms “halo” or “halogen,” by themselves or as part of anothersubstituent, mean, unless otherwise stated, a fluorine, chlorine,bromine, or iodine atom. Additionally, terms such as “haloalkyl,” aremeant to include monohaloalkyl and polyhaloalkyl. For example, the term“C₁₋₄ haloalkyl” is mean to include trifluoromethyl,2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like.

The term “aryl” means, unless otherwise stated, a polyunsaturated,typically aromatic, hydrocarbon group which can be a single ring ormultiple rings (up to three rings) which are fused together or linkedcovalently. The term “heteroaryl” refers to aryl groups (or rings) thatcontain from one to five heteroatoms selected from N, O, and S, whereinthe nitrogen and sulfur atoms are optionally oxidized, and the nitrogenatom(s) are optionally quaternized. A heteroaryl group can be attachedto the remainder of the molecule through a heteroatom. Non-limitingexamples of aryl groups include phenyl, naphthyl and biphenyl, whilenon-limiting examples of heteroaryl groups include pyridyl, pyridazinyl,pyrazinyl, pyrimindinyl, triazinyl, quinolinyl, quinoxalinyl,quinazolinyl, cinnolinyl, phthalaziniyl, benzotriazinyl, purinyl,benzimidazolyl, benzopyrazolyl, benzotriazolyl, benzisoxazolyl,isobenzofuryl, isoindolyl, indolizinyl, benzotriazinyl, thienopyridinyl,thienopyrimidinyl, pyrazolopyrimidinyl, imidazopyridines,benzothiaxolyl, benzofuranyl, benzothienyl, indolyl, quinolyl,isoquinolyl, isothiazolyl, pyrazolyl, indazolyl, pteridinyl, imidazolyl,triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiadiazolyl, pyrrolyl,thiazolyl, furyl, thienyl and the like. Substituents for each of theabove noted aryl and heteroaryl ring systems are selected from the groupof acceptable substituents described below.

For brevity, the term “aryl” when used in combination with other terms(e.g., aryloxy, arylthioxy, arylalkyl) includes both aryl and heteroarylrings as defined above. Thus, the term “arylalkyl” is meant to includethose radicals in which an aryl group is attached to an alkyl group(e.g., benzyl, phenethyl, pyridylmethyl and the like).

The above terms (e.g., “alkyl,” “aryl” and “heteroaryl”), in someembodiments, will include both substituted and unsubstituted forms ofthe indicated radical. Preferred substituents for each type of radicalare provided below. For brevity, the terms aryl and heteroaryl willrefer to substituted or unsubstituted versions as provided below, whilethe term “alkyl” and related aliphatic radicals is meant to refer tounsubstituted version, unless indicated to be substituted.

Substituents for the alkyl radicals (including those groups oftenreferred to as alkylene, alkenyl, alkynyl and cycloalkyl) can be avariety of groups selected from: -halogen, —OR′, —NR′R″, —SR′,—SiR′R″R′″, —OC(O)R′, —C(O)R′, —CO₂R′, —CONR′R″, —OC(O)NR′R″,—NR″C(O)R′, —NR′—C(O)NR″R′″, —NR″C(O)₂R′, —NH—C(NH₂)═NH, —NR′C(NH₂)═NH,—NH—C(NH₂)═NR′, —S(O)R′, —S(O)₂R′, —S(O)₂NR′R″, —NR'S(O)₂R″, —CN and—NO₂ in a number ranging from zero to (2 m′+1), where m′ is the totalnumber of carbon atoms in such radical. R′—, R″ and R′″ eachindependently refer to hydrogen, unsubstituted C₁₋₈ alkyl, unsubstitutedheteroalkyl, unsubstituted aryl, aryl substituted with 1-3 halogens,unsubstituted C₁₋₈ alkyl, C₁₋₈ alkoxy or C₁₋₈ thioalkoxy groups, orunsubstituted aryl-C₁₋₄ alkyl groups. When R′ and R″ are attached to thesame nitrogen atom, they can be combined with the nitrogen atom to forma 3-, 4-, 5-, 6-, or 7-membered ring. For example, —NR′R″ is meant toinclude 1-pyrrolidinyl and 4-morpholinyl.

Similarly, substituents for the aryl and heteroaryl groups are variedand are generally selected from: -halogen, —OR′, —OC(O)R′, —NR′R″, —SR′,—R′, —CN, —NO₂, —CO₂R′, —CONR′R″, —C(O)R′, —OC(O)NR′R″, —NR″C(O)R′,—NR″C(O)₂R′, —NR′—C(O)NR″R′″, —NH—C(NH₂)═NH, —NR′C(NH₂)═NH,—NH—C(NH₂)═NR′, —S(O)R′, —S(O)₂R′, —S(O)₂NR′R″, —NR'S(O)₂R″, —N₃,perfluoro(C₁-C₄)alkoxy, and perfluoro(C₁-C₄)alkyl, in a number rangingfrom zero to the total number of open valences on the aromatic ringsystem; and where R′, R″ and R′″ are independently selected fromhydrogen, C₁₋₈ alkyl, C₃₋₆ cycloalkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl,unsubstituted aryl and heteroaryl, (unsubstituted aryl)-C₁₋₄ alkyl, andunsubstituted aryloxy-C₁₋₄ alkyl. Other suitable substituents includeeach of the above aryl substituents attached to a ring atom by analkylene tether of from 1-4 carbon atoms.

Two of the substituents on adjacent atoms of the aryl or heteroaryl ringmay optionally be replaced with a substituent of the formula-T-C(O)—(CH₂)_(q)—U—, wherein T and U are independently —NH—, —O—, —CH₂—or a single bond, and q is an integer of from 0 to 2. Alternatively, twoof the substituents on adjacent atoms of the aryl or heteroaryl ring mayoptionally be replaced with a substituent of the formula-A-(CH₂)_(r)—B—, wherein A and B are independently —CH₂—, —O—, —NH—,—S—, —S(O)—, —S(O)₂—, —S(O)₂NR′— or a single bond, and r is an integerof from 1 to 3. One of the single bonds of the new ring so formed mayoptionally be replaced with a double bond. Alternatively, two of thesubstituents on adjacent atoms of the aryl or heteroaryl ring mayoptionally be replaced with a substituent of the formula—(CH₂)_(s)—X—(CH₂)_(t)—, where s and t are independently integers offrom 0 to 3, and X is —O—, —NR′—, —S—, —S(O)—, —S(O)₂—, or —S(O)₂NR′—.The substituent R′ in —NR′— and —S(O)₂NR′— is selected from hydrogen orunsubstituted C₁₋₆ alkyl.

As used herein, the term “heteroatom” is meant to include oxygen (O),nitrogen (N), sulfur (S) and silicon (Si).

The term “pharmaceutically acceptable salts” is meant to include saltsof the active compounds which are prepared with relatively nontoxicacids or bases, depending on the particular substituents found on thecompounds described herein. When compounds of the present inventioncontain relatively acidic functionalities, base addition salts can beobtained by contacting the neutral form of such compounds with asufficient amount of the desired base, either neat or in a suitableinert solvent. Examples of salts derived frompharmaceutically-acceptable inorganic bases include aluminum, ammonium,calcium, copper, ferric, ferrous, lithium, magnesium, manganic,manganous, potassium, sodium, zinc and the like. Salts derived frompharmaceutically-acceptable organic bases include salts of primary,secondary and tertiary amines, including substituted amines, cyclicamines, naturally-occurring amines and the like, such as arginine,betaine, caffeine, choline, N,N′-dibenzylethylenediamine, diethylamine,2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine,ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine,glucosamine, histidine, hydrabamine, isopropylamine, lysine,methylglucamine, morpholine, piperazine, piperadine, polyamine resins,procaine, purines, theobromine, triethylamine, trimethylamine,tripropylamine, tromethamine and the like. When compounds of the presentinvention contain relatively basic functionalities, acid addition saltscan be obtained by contacting the neutral form of such compounds with asufficient amount of the desired acid, either neat or in a suitableinert solvent. Examples of pharmaceutically acceptable acid additionsalts include those derived from inorganic acids like hydrochloric,hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric,monohydrogenphosphoric, dihydrogenphosphoric, sulfuric,monohydrogensulfuric, hydriodic, or phosphorous acids and the like, aswell as the salts derived from relatively nontoxic organic acids likeacetic, propionic, isobutyric, malonic, benzoic, succinic, suberic,fumaric, mandelic, phthalic, benzenesulfonic, p-tolylsulfonic, citric,tartaric, methanesulfonic, and the like. Also included are salts ofamino acids such as arginate and the like, and salts of organic acidslike glucuronic or galactunoric acids and the like (see, for example,Berge, S. M., et al, “Pharmaceutical Salts”, Journal of PharmaceuticalScience, 1977, 66, 1-19). Certain specific compounds of the presentinvention contain both basic and acidic functionalities that allow thecompounds to be converted into either base or acid addition salts.

The neutral forms of the compounds may be regenerated by contacting thesalt with a base or acid and isolating the parent compound in theconventional manner. The parent form of the compound differs from thevarious salt forms in certain physical properties, such as solubility inpolar solvents, but otherwise the salts are equivalent to the parentform of the compound for the purposes of the present invention.

In addition to salt forms, the present invention provides compoundswhich are in a prodrug form. Prodrugs of the compounds described hereinare those compounds that readily undergo chemical changes underphysiological conditions to provide the compounds of the presentinvention. Additionally, prodrugs can be converted to the compounds ofthe present invention by chemical or biochemical methods in an ex vivoenvironment. For example, prodrugs can be slowly converted to thecompounds of the present invention when placed in a transdermal patchreservoir with a suitable enzyme or chemical reagent.

Certain compounds of the present invention can exist in unsolvated formsas well as solvated forms, including hydrated forms. In general, thesolvated forms are equivalent to unsolvated forms and are intended to beencompassed within the scope of the present invention. Certain compoundsof the present invention may exist in multiple crystalline or amorphousforms. In general, all physical forms are equivalent for the usescontemplated by the present invention and are intended to be within thescope of the present invention.

Certain compounds of the present invention possess asymmetric carbonatoms (optical centers) or double bonds; the racemates, diastereomers,geometric isomers, regioisomers and individual isomers (e.g., separateenantiomers) are all intended to be encompassed within the scope of thepresent invention. The compounds of the present invention may alsocontain unnatural proportions of atomic isotopes at one or more of theatoms that constitute such compounds. For example, the compounds may beradiolabeled with radioactive isotopes, such as for example tritium(³H), iodine-125 (¹²⁵I) or carbon-14 (¹⁴C). All isotopic variations ofthe compounds of the present invention, whether radioactive or not, areintended to be encompassed within the scope of the present invention.

II. General

The present invention derives from the discovery that compounds offormula I (as well as the subgeneric formulae Ia, Ia1, Ia2, II, III andIV) act as potent antagonists of the CCR1 receptor. The compounds havein vivo anti-inflammatory activity. Accordingly, the compounds providedherein are useful in pharmaceutical compositions, methods for thetreatment of CCR1-mediated diseases, and as controls in assays for theidentification of competitive CCR1 antagonists.

III. Compounds

In one aspect, the present invention provides compounds having theformula:

and salts, rotamers and optical isomers thereof.

In formula I, the subscript n is an integer of from 0 to 3; each R^(1a)and R^(1b) is a member independently selected from the group consistingof H, C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₃₋₆ cycloalkyl, —COR^(a), —CO₂R^(a),—CONR^(a)R^(b), —NR^(a)R^(b), —NR^(a)CO₂R^(b), —OR^(a), —X¹COR^(a),—X¹CO₂R^(a), —X¹CONR^(a)R^(b), —X¹NR^(a)COR^(b), —X¹NR^(a)R^(b), and—X¹OR^(a), wherein X¹ is a member selected from the group consisting ofC₁₋₄ alkylene, and each R^(a) and R^(b) is independently selected fromthe group consisting of hydrogen, C₁₋₈ alkyl, C₁₋₈ haloalkyl, and C₃₋₆cycloalkyl, and optionally two R^(1a) groups on adjacent carbon atomsare joined to form a 5-, 6- or 7-membered carbocyclic or heterocyclicring; each of R^(2a) and R^(2b) is a member independently selected fromthe group consisting of H, hydroxyl, C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₁₋₈alkoxy, C₁₋₄ alkoxy-C₁₋₄ alkyl, C₁₋₈ hydroxyalkyl, C₁₋₄ alkoxy-C₁₋₄alkoxy, C₃₋₆ cycloalkyl, C₃₋₆ cycloalkyl-C₁₋₄ alkyl, 3- to 7-memberedheterocycloalkyl, 3- to 7-membered heterocycloalkyl-C₁₋₄ alkyl,—X¹CO₂R^(a), —X¹CONR^(a)R^(b), —X¹NR^(a)COR^(b), —X¹NR^(a)R^(b), whereinX¹, R^(a) and R^(b) are defined above.

The symbol Ar¹ represents a six- or ten-membered monocyclic or fusedbicyclic aryl ring, or a five- to ten-membered monocyclic or fusedbicyclic heteroaryl ring; each of which is substituted with from one tofive substituents, R³, R^(3a), R^(3b), R⁴ and R^(4a) which areindependently selected from the group consisting of H, halogen, —OR^(c),—OC(O)R^(c), —NR^(c)R^(d), —SR^(c), —R^(e), —CN, —NO₂, —CO₂R^(c),—CONR^(c)R^(d), —C(O)R^(c), —OC(O)NR^(c)R^(d), —NR^(d)C(O)R^(c),—NR^(d)C(O)₂R^(e), —NR^(c)—C(O)NR^(c)R^(d), —NH—C(NH₂)═NH,—NR^(e)C(NH₂)═NH, —NH—C(NH₂)═NR^(e), —NH—C(NHR^(e))═NH, —S(O)R^(e),—S(O)₂R^(e), —NR^(c)S(O)₂R^(e), —S(O)₂NR^(c)R^(d), —N₃, —X²OR^(c),—O—X²OR^(c), —X²OC(O)R^(c), —X²NR^(c)R^(d), —O—X²NR^(c)R^(d), —X²SR^(c),—X²CN, —X²NO₂, —X²CO₂R^(c), —O—X²CO₂R^(c), —X²CONR^(c)R^(d),—O—X²CONR^(c)R^(d), —X²C(O)R^(c), —X²OC(O)NR^(c)R^(d),—X²NR^(d)C(O)R^(c), —X²NR^(d)C(O)₂R^(e), —X²NR^(c)C(O)NR^(c)R^(d),—X²NH—C(NH₂)═NH, —X²NR^(e)C(NH₂)═NH, —X²NH—C(NH₂)═NR^(e),—X²NH—C(NHR^(e))═NH, —X²S(O)R^(e), —X²S(O)₂R^(e), —X²NR^(c)S(O)₂R^(e),—X²S(O)₂NR^(c)R^(d), —X²N₃, —NR^(d)—X₂OR^(c), —NR^(d)—X²NR^(c)R^(d),—NR^(d)—X²CO₂R^(c), and —NR^(d)—X²CONR^(c)R^(d), wherein each X² is amember independently selected from the group consisting of C₁₋₄alkylene, and each R^(c) and R^(d) is independently selected fromhydrogen, C₁₋₈ alkyl, C₁₋₈ hydroxyalkyl, C₁₋₈ haloalkyl and C₃₋₆cycloalkyl, or optionally R^(e) and R^(d) when attached to the samenitrogen atom can be combined with the nitrogen atom to form a five orsix-membered ring having from 0 to 2 additional heteroatoms as ringmembers; and each R^(e) is independently selected from the groupconsisting of C₁₋₈ alkyl, C₁₋₈ hydroxyalkyl, C₁₋₈ haloalkyl and C₃₋₆cycloalkyl.

The symbol Ar² represents a six- or ten-membered monocyclic or fusedbicyclic aryl ring, or a five- to ten-membered monocyclic or fusedbicyclic heteroaryl ring; each of which is substituted with from one tofive substituents, R⁵, R⁶, R⁷, R⁸ and R⁹, independently selected fromthe group consisting of H, halogen, —OR^(f), —OC(O)R^(f), —NR^(f)R^(g),—SR, —R^(h), —CN, —NO₂, —CO₂R^(f), —CONR^(f)R^(g), —C(O)R^(f),—OC(O)NR^(f)R^(g), —NR^(g)C(O)R^(f), —NR^(g)C(O)₂R^(h),—NR^(f)—C(O)NR^(f)R^(g), —NH—C(NH₂)═NH, —NR^(h)C(NH₂)═NH,—NH—C(NH₂)═NR^(h), —NH—C(NHR^(h))═NH, —S(O)R^(h), —S(O)₂R^(h),—NR^(f)S(O)₂R^(h), —S(O)₂NR^(f)R^(g), —NR^(f)S(O)₂NR^(f)R^(g), —N₃,—X³OR^(f), —X³OC(O)R^(f), —X³NR^(f)R^(g), —X³SR′, —X³CN, —X³NO₂,—X³CO₂R^(f), —X³CONR^(f)R^(g), —X³C(O)R^(f), —X³OC(O)NR^(f)R^(g),—X³NR^(g)C(O)R^(f), —X³NR^(g)C(O)₂R^(h), —X³NR^(f)—C(O)NR^(f)R^(g),—X³NH—C(NH₂)═NH, —X³NR^(h)C(NH₂)═NH, —X³NH—C(NH₂)═NR^(h),—X³NH—C(NHR^(h))—NH, —X³S(O)R^(h), —X³S(O)₂R^(h), —X³NR^(f)S(O)₂R^(h),—X³S(O)₂NR^(f)R^(g), —Y, —X³Y, —S(O)₂Y, —C(O)Y, —X³N₃, —O—X³OR^(f),—O—X³NR^(f)R^(g), —O—X³CO₂R^(f), —O—X³CONR^(f)R^(g), —NR^(g)—X³OR^(f),—NR^(g)—X³NR^(f)R^(g), —NR^(g)—X³CO₂R^(f), and —NR^(g)—X³CONR^(f)R^(g),wherein Y is a five or six-membered aryl, heteroaryl or heterocyclicring, optionally substituted with from one to three substitutentsselected from the group consisting of halogen, —OR, —OC(O)R^(f),—NR^(f)R^(g), —SR^(f), —CN, —NO₂, —CO₂R^(g), —CONR^(f)R^(g), —C(O)R′,—NR^(g)C(O)R^(h), —NR^(g)C(O)₂R^(h), —S(O)R^(h), —S(O)₂R^(h),—NR^(f)S(O)₂R^(h), —S(O)₂NR^(f)R^(g), —X³OR′, —X³SR′, —X³CN, —X³NO₂,—X³CO₂R^(f), —X³CONR^(f)R^(g), —X³C(O)R^(f), —X³OC(O)NR^(f)R^(g),—X³NR^(g)C(O)R^(f), —X³NR^(g)C(O)₂R^(h), —X³NR^(f)—C(O)NR^(f)R^(g),—X³OC(O)R^(f), —X³S(O)R^(h), —X³S(O)₂R^(h), —X³NR^(f)R^(g),—X³NR^(f)S(O)₂R^(h), —X³S(O)₂NR^(f)R^(g), —O—X³OR^(f), —O—X³NR^(f)R^(g),—O—X³CO₂R^(f), —O—X³CONR^(f)R^(g), —NR^(g), —X³OR^(f),—NR^(g)—X³NR^(f)R^(g), —NR^(g)—X³CO₂R^(f), and —NR^(g)—X³CONR^(f)R^(g)and wherein each X³ is independently selected from the group consistingof C₁₋₄ alkylene, and each R^(f) and R^(g) is independently selectedfrom hydrogen, C₁₋₈ alkyl, C₁₋₈ hydroxyalkyl, C₁₋₈ haloalkyl and C₃₋₆cycloalkyl, or when attached to the same nitrogen atom can be combinedwith the nitrogen atom to form a five or six-membered ring having from 0to 2 additional heteroatoms as ring members, and each R^(h) isindependently selected from the group consisting of C₁₋₈ alkyl, C₂₋₈alkenyl, C₂₋₈ alkynyl, C₁₋₈ hydroxyalkyl, C₁₋₈ haloalkyl and C₃₋₆cycloalkyl; or when two of R⁵, R⁶, R⁷, R⁸ and R⁹, are attached toadjacent ring vertices of Ar², are optionally combined to form a five orsix membered ring having zero, one or two heteroatoms selected from Oand N as ring members.

In some embodiments, the compounds of formula I are those in which Ar¹is selected from the group consisting of phenyl, naphthyl, pyridyl,pyrazinyl, pyridazinyl, pyrimidinyl, triazinyl, quinolinyl, quinoxalinyland purinyl, each of which is optionally substituted with R³, R^(3a),R^(3b), R⁴ and R^(4a).

In other embodiments, the compounds of formula I are those in which Ar¹is selected from the group consisting of phenyl, naphthyl and pyridyl,each of which is optionally substituted with R³, R^(3a), R^(3b), R⁴ andR^(4a).

In still other embodiments, the compounds of formula I are those inwhich Ar² is selected from the group consisting of phenyl, pyrazolyl,imidazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl,oxathiadiazolyl, pyrrolyl, thiazolyl, isothiazolyl, benzimidazolyl,benzoxazolyl, benzopyrazolyl, benzotriazolyl, pyrazolo[3,4-b]pyridine,pyrazolo[3,4-d]pyrimidine, imidazo[4,5-b]pyri dine,imidazo[1,5-α]pyridine, and pyrrolo[2,3-b]pyridine, each of which isoptionally substituted with R⁵, R⁶ and R⁷.

In yet other embodiments, the compounds of formula I are those in whichAr² is selected from the group consisting of pyrazolyl, imidazolyl andtriazolyl, each of which is substituted with R⁵, R⁶ and R⁷.

In certain embodiments, the compounds of formula I are those in whichAr¹ is selected from the group consisting of phenyl, naphthyl andpyridyl, each of which is substituted with from one to fivesubstituents, R³, R^(3a), R^(3b), R⁴ and R^(4a); and Ar² is selectedfrom the group consisting of pyrazolyl, imidazolyl and triazolyl, eachof which is substituted with R¹, R⁶ and R⁷.

In selected embodiments, the compounds of formula I are those in whichAr¹ is phenyl, which is substituted with from one to five substituents,R³, R^(3a), R^(3b), R⁴ and R^(4a), and Ar² is selected from the groupconsisting of pyrazolyl, imidazolyl, benzimidazolyl, benzopyrazolyl,pyrazolo[3,4-b]pyridine, pyrazolo[3,4-d]pyrimidine,imidazo[4,5-b]pyridine, imidazo[1,5-a]pyridine, andpyrrolo[2,3-b]pyridine, each of which is optionally substituted with R⁵,R⁶ and R⁷.

Still other embodiments of the invention are the compounds of formulaeIa, Ia1, Ia2, II, III and IV.

Accordingly, in some embodiments, the compounds are those of formula Ia:

or a pharmaceutically acceptable salt, rotamer or optical isomerthereof, wherein R³ and R⁴ are independently selected from the groupconsisting of H, halogen, —R^(e), —CN, and —SO₂R^(e); and the groupsR^(1a), R^(2a) and Ar² have the meanings provided with reference toformula I above, or the other embodiments provided.

In still other embodiments of formula I or Ia, Ar² is a heteroarylgroup; in other embodiments, Ar² is a heteroaryl group, optionallysubstituted and attached to the remainder of the molecule through anitrogen atom ring vertex; and in still other embodiments, Ar² has theformula:

wherein R⁵, R⁶, and R⁷ are independently selected from the groupconsisting of H, halogen, —R^(h), —CN, —SO₂R^(h), —CO₂R^(f),—CONR^(f)R^(g), and Y, wherein —R^(h), R^(f), R^(g), and Y have themeanings provided above with respect to formula I.

In one group of selected embodiments, the compounds have the formula:

or a pharmaceutically acceptable salt, rotamer or optical isomerthereof, wherein R⁴ is selected from the group consisting of F and Cl;and the groups R^(1a), R^(2a), R³ and Ar² have the meanings providedwith reference to formula I or Ia above, or the other embodimentsprovided.

In another group of selected embodiments, the compounds have theformula:

or a pharmaceutically acceptable salt, rotamer or optical isomerthereof, wherein R³ is selected from the group consisting of H, halogen,C₁₋₈ alkyl, C₁₋₈ haloalkyl and C₁₋₈ alkoxy; R^(1a) and R^(2a) areindependently selected from the group consisting of H, C₁₋₈ alkyl, C₁₋₈haloalkyl, C₁₋₄ alkoxy and C₁₋₈ hydroxyalkyl; and Ar² has the meaningprovided with reference to formula I or Ia above, or the embodimentsprovided.

In yet another group of selected embodiments, the compounds have theformula:

or a pharmaceutically acceptable salt, rotamer or optical isomerthereof, wherein R^(1a) and R^(2a) are independently selected from thegroup consisting of H, C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₁₋₈ alkoxy and C₁₋₈hydroxyalkyl; and R⁵, R⁶, and R⁷ are independently selected from thegroup consisting of H, halogen, —R^(h), —CN, —SO₂R^(h), —CO₂R^(f),—CONR^(f)R^(g), and Y; wherein —R^(h), R^(f), R^(g), and Y have themeanings provided above with respect to formula I.

In still another group of selected embodiments, the compounds have theformula:

or a pharmaceutically acceptable salt, rotamer or optical isomerthereof, wherein R^(1a) and R^(2a) are independently selected from thegroup consisting of H, C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₁₋₈ alkoxy and C₁₋₈hydroxyalkyl; and R⁵, R⁶, and R⁷ are independently selected from thegroup consisting of H, halogen, —R^(h), —CN, —SO₂R^(h), —CO₂R^(f),—CONR^(f)R^(g), and Y; wherein —R^(h), R^(f), R^(g), and Y have themeanings provided above with respect to formula I.

In another group of selected embodiments, the compounds have theformula:

or a pharmaceutically acceptable salt, rotamer or optical isomerthereof, wherein R^(1a) and R^(2a) are each independently selected fromthe group consisting of H, C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₁₋₈ alkoxy, andC₁₋₈ hydroxyalkyl; and R⁵ and R⁷ are each independently selected fromthe group consisting of H, halogen, —R^(h), —CN, —SO₂R^(h), —CO₂R^(f),—CONR^(f)R^(g), and Y; wherein —R^(h), R^(f), R^(g), and Y have themeanings provided above with respect to formula I.

For any of the embodiments above, when Y is present, selectedembodiments are those in which Y is selected from the group consistingof pyridyl, pyrimidinyl, imidazolyl, oxazolyl, oxadiazolyl, triazolyl,thiazolyl, imidazolinyl and pyrazolyl.

Specific compounds of particular interest are those provided in Table 1,along with their pharmaceutically acceptable salts, hydrates orN-oxides, rotamers, and stereoisomers thereof.

Preparation of Compounds

As provided in the examples below, the compounds and intermediates ofthe present invention can be prepared by one of skill in the art in acomponent assembly manner.

IV. Pharmaceutical Compositions

In addition to the compounds provided above, compositions for modulatingCCR1 activity in humans and animals will typically contain apharmaceutical carrier or diluent.

The term “composition” as used herein is intended to encompass a productcomprising the specified ingredients in the specified amounts, as wellas any product which results, directly or indirectly, from combinationof the specified ingredients in the specified amounts. By“pharmaceutically acceptable” it is meant the carrier, diluent orexcipient must be compatible with the other ingredients of theformulation and not deleterious to the recipient thereof.

The pharmaceutical compositions for the administration of the compoundsof this invention may conveniently be presented in unit dosage form andmay be prepared by any of the methods well known in the art of pharmacyand drug delivery. All methods include the step of bringing the activeingredient into association with the carrier which constitutes one ormore accessory ingredients. In general, the pharmaceutical compositionsare prepared by uniformly and intimately bringing the active ingredientinto association with a liquid carrier or a finely divided solid carrieror both, and then, if necessary, shaping the product into the desiredformulation. In the pharmaceutical composition the active objectcompound is included in an amount sufficient to produce the desiredeffect upon the process or condition of diseases.

The pharmaceutical compositions containing the active ingredient may bein a form suitable for oral use, for example, as tablets, troches,lozenges, aqueous or oily suspensions, dispersible powders or granules,emulsions and self emulsifications as described in U.S. PatentApplication 2002-0012680, hard or soft capsules, syrups, elixirs,solutions, buccal patch, oral gel, chewing gum, chewable tablets,effervescent powder and effervescent tablets. Compositions intended fororal use may be prepared according to any method known to the art forthe manufacture of pharmaceutical compositions and such compositions maycontain one or more agents selected from the group consisting ofsweetening agents, flavoring agents, coloring agents, antioxidants andpreserving agents in order to provide pharmaceutically elegant andpalatable preparations. Tablets contain the active ingredient inadmixture with non-toxic pharmaceutically acceptable excipients whichare suitable for the manufacture of tablets. These excipients may be forexample, inert diluents, such as cellulose, silicon dioxide, aluminumoxide, calcium carbonate, sodium carbonate, glucose, mannitol, sorbitol,lactose, calcium phosphate or sodium phosphate; granulating anddisintegrating agents, for example, corn starch, or alginic acid;binding agents, for example PVP, cellulose, PEG, starch, gelatin oracacia, and lubricating agents, for example magnesium stearate, stearicacid or talc. The tablets may be uncoated or they may be coated,enterically or otherwise, by known techniques to delay disintegrationand absorption in the gastrointestinal tract and thereby provide asustained action over a longer period. For example, a time delaymaterial such as glyceryl monostearate or glyceryl distearate may beemployed. They may also be coated by the techniques described in theU.S. Pat. Nos. 4,256,108; 4,166,452; and U.S. Pat. No. 4,265,874 to formosmotic therapeutic tablets for control release.

Formulations for oral use may also be presented as hard gelatin capsuleswherein the active ingredient is mixed with an inert solid diluent, forexample, calcium carbonate, calcium phosphate or kaolin, or as softgelatin capsules wherein the active ingredient is mixed with water or anoil medium, for example peanut oil, liquid paraffin, or olive oil.Additionally, emulsions can be prepared with a non-water miscibleingredient such as oils and stabilized with surfactants such asmono-diglycerides, PEG esters and the like.

Aqueous suspensions contain the active materials in admixture withexcipients suitable for the manufacture of aqueous suspensions. Suchexcipients are suspending agents, for example sodiumcarboxymethylcellulose, methylcellulose, hydroxy-propylmethylcellulose,sodium alginate, polyvinyl-pyrrolidone, gum tragacanth and gum acacia;dispersing or wetting agents may be a naturally-occurring phosphatide,for example lecithin, or condensation products of an alkylene oxide withfatty acids, for example polyoxy-ethylene stearate, or condensationproducts of ethylene oxide with long chain aliphatic alcohols, forexample heptadecaethyleneoxycetanol, or condensation products ofethylene oxide with partial esters derived from fatty acids and ahexitol such as polyoxyethylene sorbitol monooleate, or condensationproducts of ethylene oxide with partial esters derived from fatty acidsand hexitol anhydrides, for example polyethylene sorbitan monooleate.The aqueous suspensions may also contain one or more preservatives, forexample ethyl, or n-propyl, p-hydroxybenzoate, one or more coloringagents, one or more flavoring agents, and one or more sweetening agents,such as sucrose or saccharin.

Oily suspensions may be formulated by suspending the active ingredientin a vegetable oil, for example arachis oil, olive oil, sesame oil orcoconut oil, or in a mineral oil such as liquid paraffin. The oilysuspensions may contain a thickening agent, for example beeswax, hardparaffin or cetyl alcohol. Sweetening agents such as those set forthabove, and flavoring agents may be added to provide a palatable oralpreparation. These compositions may be preserved by the addition of ananti-oxidant such as ascorbic acid.

Dispersible powders and granules suitable for preparation of an aqueoussuspension by the addition of water provide the active ingredient inadmixture with a dispersing or wetting agent, suspending agent and oneor more preservatives. Suitable dispersing or wetting agents andsuspending agents are exemplified by those already mentioned above.Additional excipients, for example sweetening, flavoring and coloringagents, may also be present.

The pharmaceutical compositions of the invention may also be in the formof oil-in-water emulsions. The oily phase may be a vegetable oil, forexample olive oil or arachis oil, or a mineral oil, for example liquidparaffin or mixtures of these. Suitable emulsifying agents may benaturally-occurring gums, for example gum acacia or gum tragacanth,naturally-occurring phosphatides, for example soy bean, lecithin, andesters or partial esters derived from fatty acids and hexitolanhydrides, for example sorbitan monooleate, and condensation productsof the said partial esters with ethylene oxide, for examplepolyoxyethylene sorbitan monooleate. The emulsions may also containsweetening and flavoring agents.

Syrups and elixirs may be formulated with sweetening agents, for exampleglycerol, propylene glycol, sorbitol or sucrose. Such formulations mayalso contain a demulcent, a preservative and flavoring and coloringagents. Oral solutions can be prepared in combination with, for example,cyclodextrin, PEG and surfactants.

The pharmaceutical compositions may be in the form of a sterileinjectable aqueous or oleagenous suspension. This suspension may beformulated according to the known art using those suitable dispersing orwetting agents and suspending agents which have been mentioned above.The sterile injectable preparation may also be a sterile injectablesolution or suspension in a non-toxic parenterally-acceptable diluent orsolvent, for example as a solution in 1,3-butane diol. Among theacceptable vehicles and solvents that may be employed are water,Ringer's solution and isotonic sodium chloride solution. In addition,sterile, fixed oils are conventionally employed as a solvent orsuspending medium. For this purpose any bland fixed oil may be employedincluding synthetic mono- or diglycerides. In addition, fatty acids suchas oleic acid find use in the preparation of injectables.

The compounds of the present invention may also be administered in theform of suppositories for rectal administration of the drug. Thesecompositions can be prepared by mixing the drug with a suitablenon-irritating excipient which is solid at ordinary temperatures butliquid at the rectal temperature and will therefore melt in the rectumto release the drug. Such materials include cocoa butter andpolyethylene glycols. Additionally, the compounds can be administeredvia ocular delivery by means of solutions or ointments. Still further,transdermal delivery of the subject compounds can be accomplished bymeans of iontophoretic patches and the like. For topical use, creams,ointments, jellies, solutions or suspensions, etc., containing thecompounds of the present invention are employed. As used herein, topicalapplication is also meant to include the use of mouth washes andgargles.

The compounds of this invention may also be coupled to a carrier that isa suitable polymers as targetable drug carriers. Such polymers caninclude polyvinylpyrrolidone, pyran copolymer,polyhydroxy-propyl-methacrylamide-phenol,polyhydroxyethyl-aspartamide-phenol, or polyethyleneoxide-polylysinesubstituted with palmitoyl residues. Furthermore, the compounds of theinvention may be coupled to a carrier that is a class of biodegradablepolymers useful in achieving controlled release of a drug, for examplepolylactic acid, polyglycolic acid, copolymers of polylactic andpolyglycolic acid, polyepsilon caprolactone, polyhydroxy butyric acid,polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates andcross linked or amphipathic block copolymers of hydrogels. Polymers andsemipermeable polymer matrices may be formed into shaped articles, suchas valves, stents, tubing, prostheses and the like. In one embodiment ofthe invention, the compound of the invention is coupled to a polymer orsemipermeable polymer matrix that is formed as a stent or stent-graftdevice.

V. Methods of Treating Diseases Modulated by CCR1

In yet another aspect, the present invention provides methods oftreating CCR1-mediated conditions or diseases by administering to asubject having such a disease or condition, a therapeutically effectiveamount of a compound of formula I above. The “subject” is defined hereinto include animals such as mammals, including, but not limited to,primates (e.g., humans), cows, sheep, goats, horses, dogs, cats,rabbits, rats, mice and the like.

CCR1 provides a target for interfering with or promoting specificaspects of immune cell functions, or more generally, with functionsassociated with CCR1 expression on a wide range of cell types in amammal, such as a human. Compounds that inhibit CCR1, are particularlyuseful for modulating monocyte, macrophage, lymphocyte, granulocyte, NKcell, mast cells, dendritic cell, neutrophils, and certain immunederived cell (for example, osteoclasts) function for therapeuticpurposes. Accordingly, the present invention is directed to compoundswhich are useful in the prevention and/or treatment of a wide variety ofinflammatory and immunoregulatory disorders and diseases (see Saeki, etal., Current Pharmaceutical Design 9:1201-1208 (2003)).

For example, an instant compound that inhibits one or more functions ofCCR1 may be administered to inhibit (i.e., reduce or prevent)inflammation or cellular infiltration associated with an immunedisorder. As a result, one or more inflammatory processes, such asleukocyte emigration or infiltration, chemotaxis, exocytosis (e.g., ofenzymes, histamine) or inflammatory mediator release, can be inhibited.For example, monocyte infiltration to an inflammatory site (e.g., anaffected joint in arthritis, or into the CNS in MS) can be inhibitedaccording to the present method.

Similarly, an instant compound that promotes one or more functions ofCCR1 is administered to stimulate (induce or enhance) an inflammatoryresponse, such as leukocyte emigration, chemotaxis, exocytosis (e.g., ofenzymes, histamine) or inflammatory mediator release, resulting in thebeneficial stimulation of inflammatory processes. For example, monocytescan be recruited to combat bacterial infections.

Diseases and conditions associated with inflammation, immune disordersand infection can be treated using the method of the present invention.In a preferred embodiment, the disease or condition is one in which theactions of immune cells such monocyte, macrophage, lymphocyte,granulocyte, NK cell, mast cell, dendritic cell, or certain immunederived cell (for example, osteoclasts) are to be inhibited or promoted,in order to modulate the inflammatory or autoimmune response.

In one group of embodiments, diseases or conditions, including chronicdiseases, of humans or other species can treated with modulators of CCR1function. These diseases or conditions include: (1) allergic diseasessuch as systemic anaphylaxis or hypersensitivity responses, drugallergies, insect sting allergies and food allergies, (2) inflammatorybowel diseases, such as Crohn's disease, ulcerative colitis, ileitis andenteritis, (3) vaginitis, (4) psoriasis and inflammatory dermatoses suchas dermatitis, eczema, atopic dermatitis, allergic contact dermatitis,urticaria and pruritus, (5) vasculitis, (6) spondyloarthropathies, (7)scleroderma, (8) asthma and respiratory allergic diseases such asallergic asthma, allergic rhinitis, hypersensitivity lung diseases andthe like, (9) autoimmune diseases, such as fibromyalagia, scleroderma,ankylosing spondylitis, juvenile RA, Still's disease, polyarticularjuvenile RA, pauciarticular juvenile RA, polymyalgia rheumatica,rheumatoid arthritis, psoriatic arthritis, osteoarthritis, polyarticulararthritis, multiple sclerosis, systemic lupus erythematosus, type Idiabetes, type II diabetes, glomerulonephritis, and the like, (10) graftrejection (including allograft rejection and graft-v-host disease), and(11) other diseases in which undesired inflammatory responses or immunedisorders are to be inhibited, such as cardiovascular disease includingatherosclerosis and restenosis, myositis, neurodegenerative diseases(e.g., Alzheimer's disease), encephalitis, meningitis, hepatitis,nephritis, sepsis, sarcoidosis, allergic conjunctivitis, otitis, chronicobstructive pulmonary disease, sinusitis, Behcet's syndrome and gout,(12) osteoporosis and other disorders of the bone (13) immune mediatedfood allergies such as Celiac disease and (14) radiation-inducedpulmonary disease (RIPD). See eg Yang et al, Am J Respir Cell Mol Biol.45(1):127-35 (2011).

In another group of embodiments, diseases or conditions can be treatedwith modulators of CCR1 function. Examples of diseases to be treatedwith modulators of CCR1 function include cancers such as multiplemyeloma and related osteolytic bone disease, cardiovascular diseases,diseases in which angiogenesis or neovascularization play a role(neoplastic diseases, retinopathy and macular degeneration), infectiousdiseases (viral infections, e.g., HIV and RSV infection, and bacterialinfections) and immunosuppressive diseases such as organ transplantconditions and skin transplant conditions. The term “organ transplantconditions” is meant to include bone marrow transplant conditions andsolid organ (e.g., kidney, liver, lung, heart, pancreas or combinationthereof) transplant conditions.

The compounds of the present invention are accordingly useful in theprevention and treatment of a wide variety of inflammatory andimmunoregulatory disorders and diseases.

Depending on the disease to be treated and the subject's condition, thecompounds of the present invention may be administered by oral,parenteral (e.g., intramuscular, intraperitoneal, intravenous, ICV,intracisternal injection or infusion, subcutaneous injection, orimplant), by implantation (e.g., as when the compound is coupled to astent device), by inhalation spray, nasal, vaginal, rectal, sublingual,or topical routes of administration and may be formulated, alone ortogether, in suitable dosage unit formulations containing conventionalnon-toxic pharmaceutically acceptable carriers, adjuvants and vehiclesappropriate for each route of administration.

In the treatment or prevention of conditions which require chemokinereceptor modulation an appropriate dosage level will generally be about0.001 to 100 mg per kg patient body weight per day which can beadministered in single or multiple doses. Preferably, the dosage levelwill be about 0.01 to about 25 mg/kg per day; more preferably about 0.05to about 10 mg/kg per day. A suitable dosage level may be about 0.01 to25 mg/kg per day, about 0.05 to 10 mg/kg per day, or about 0.1 to 5mg/kg per day. Within this range the dosage may be 0.005 to 0.05, 0.05to 0.5 or 0.5 to 5.0 mg/kg per day. For oral administration, thecompositions are preferably provided in the form of tablets containing1.0 to 1000 milligrams of the active ingredient, particularly 1.0, 5.0,10.0, 15.0, 20.0, 25.0, 50.0, 75.0, 100.0, 150.0, 200.0, 250.0, 300.0,400.0, 500.0, 600.0, 750.0, 800.0, 900.0, and 1000.0 milligrams of theactive ingredient for the symptomatic adjustment of the dosage to thepatient to be treated. The compounds may be administered on a regimen of1 to 4 times per day, preferably once or twice per day.

It will be understood, however, that the specific dose level andfrequency of dosage for any particular patient may be varied and willdepend upon a variety of factors including the activity of the specificcompound employed, the metabolic stability and length of action of thatcompound, the age, body weight, hereditary characteristics, generalhealth, sex and diet of the subject, as well as the mode and time ofadministration, rate of excretion, drug combination, and the severity ofthe particular condition for the subject undergoing therapy.

Diseases and conditions associated with inflammation, immune disorder,infection and cancer can be treated or prevented with the presentcompounds, compositions, and methods.

The compounds and compositions of the present invention can be combinedwith other compounds and compositions having related utilities toprevent and treat the condition or disease of interest, such asinflammatory or autoimmune disorders, conditions and diseases, includinginflammatory bowel disease, rheumatoid arthritis, osteoarthritis,psoriatic arthritis, polyarticular arthritis, multiple sclerosis,allergic diseases, psoriasis, atopic dermatitis and asthma, and thosepathologies noted above.

For example, in the treatment or prevention of inflammation orautoimmunity or for example arthritis associated bone loss, the presentcompounds and compositions may be used in conjunction with ananti-inflammatory or analgesic agent such as an opiate agonist, alipoxygenase inhibitor, such as an inhibitor of 5-lipoxygenase, acyclooxygenase inhibitor, such as a cyclooxygenase-2 inhibitor, aninterleukin inhibitor, such as an interleukin-1 inhibitor, an NMDAantagonist, an inhibitor of nitric oxide or an inhibitor of thesynthesis of nitric oxide, a non steroidal anti-inflammatory agent, or acytokine-suppressing anti-inflammatory agent, for example with acompound such as acetaminophen, aspirin, codeine, fentanyl, ibuprofen,indomethacin, ketorolac, morphine, naproxen, phenacetin, piroxicam, asteroidal analgesic, sufentanyl, sunlindac, tenidap, and the like.Similarly, the instant compounds and compositions may be administeredwith an analgesic listed above; a potentiator such as caffeine, an H2antagonist (e.g., ranitidine), simethicone, aluminum or magnesiumhydroxide; a decongestant such as phenylephrine, phenylpropanolamine,pseudoephedrine, oxymetazoline, ephinephrine, naphazoline,xylometazoline, propylhexedrine, or levo desoxy ephedrine; anantitussive such as codeine, hydrocodone, caramiphen, carbetapentane, ordextromethorphan; a diuretic; and a sedating or non sedatingantihistamine.

Likewise, compounds and compositions of the present invention may beused in combination with other drugs that are used in the treatment,prevention, suppression or amelioration of the diseases or conditionsfor which compounds and compositions of the present invention areuseful. Such other drugs may be administered, by a route and in anamount commonly used therefor, contemporaneously or sequentially with acompound or composition of the present invention. When a compound orcomposition of the present invention is used contemporaneously with oneor more other drugs, a pharmaceutical composition containing such otherdrugs in addition to the compound or composition of the presentinvention is preferred. Accordingly, the pharmaceutical compositions ofthe present invention include those that also contain one or more otheractive ingredients or therapeutic agents, in addition to a compound orcomposition of the present invention. Examples of other therapeuticagents that may be combined with a compound or composition of thepresent invention, either administered separately or in the samepharmaceutical compositions, include, but are not limited to: (a) VLA-4antagonists, (b) corticosteroids, such as beclomethasone,methylprednisolone, betamethasone, prednisone, prenisolone,dexamethasone, fluticasone, hydrocortisone, budesonide, triamcinolone,salmeterol, salmeterol, salbutamol, formeterol; (c) immunosuppressantssuch as cyclosporine (cyclosporine A, Sandimmune®, Neoral®), tacrolirnus(FK-506, Prograf®), rapamycin (sirolimus, Rapamune®) and other FK-506type immunosuppressants, and mycophenolate, e.g., mycophenolate mofetil(CellCept®); (d) antihistamines (H1-histamine antagonists) such asbromopheniramine, chlorpheniramine, dexchloipheniramine, triprolidine,clemastine, diphenhydramine, diphenylpyraline, tripelennamine,hydroxyzine, methdilazine, promethazine, trimeprazine, azatadine,cyproheptadine, antazoline, pheniramine pyrilamine, astemizole,terfenadine, loratadine, cetirizine, fexofenadine,descarboethoxyloratadine, and the like; (e) non steroidal antiasthmatics (e.g., terbutaline, metaproterenol, fenoterol, isoetharine,albuterol, bitolterol and pirbuterol), theophylline, cromolyn sodium,atropine, ipratropium bromide, leukotriene antagonists (e.g.,zafmlukast, montelukast, pranlukast, iralukast, pobilukast andSKB-106,203), leukotriene biosynthesis inhibitors (zileuton, BAY-1005);(f) non steroidal anti-inflammatory agents (NSAIDs) such as propionicacid derivatives (e.g., alminoprofen, benoxaprofen, bucloxic acid,carprofen, fenbufen, fenoprofen, fluprofen, flurbiprofen, ibuprofen,indoprofen, ketoprofen, rniroprofen, naproxen, oxaprozin, pirprofen,pranoprofen, suprofen, tiaprofenic acid and tioxaprofen), acetic acidderivatives (e.g., indomethacin, acemetacin, alclofenac, clidanac,diclofenac, fenclofenac, fenclozic acid, fentiazac, furofenac, ibufenac,isoxepac, oxpinac, sulindac, tiopinac, tolmetin, zidometacin andzomepirac), fenamic acid derivatives (e.g., flufenamic acid,meclofenamic acid, mefenamic acid, niflumic acid and tolfenamic acid),biphenylcarboxylic acid derivatives (e.g., diflunisal and flufenisal),oxicams (e.g., isoxicam, piroxicam, sudoxicam and tenoxican),salicylates (e.g., acetyl salicylic acid and sulfasalazine) and thepyrazolones (e.g., apazone, bezpiperylon, feprazone, mofebutazone,oxyphenbutazone and phenylbutazone); (g) cyclooxygenase-2 (COX-2)inhibitors such as celecoxib (Celebrex®) and rofecoxib (Vioxx®); (h)inhibitors of phosphodiesterase type IV (PDE IV); (i) gold compoundssuch as auranofin and aurothioglucose, (j) peptide and antibodymodulators of immune regulatory molecules such as etanercept (Enbrel®),infliximab (Remicade®), adalimumab (Humira®), certolizumab pegol(Cimzia®), Abatacept (Orencia®), and golimumab (Simponi®), (k) antibodytherapies such as orthoclone (OKT3), daclizumab (Zenapax®), basiliximab(Simulect®) and infliximab (Remicade®), (1) other antagonists of thechemokine receptors, especially CCR5, CXCR2, CXCR3, CCR2, CCR3, CCR4,CCR7, CX₃CR1 and CXCR6; (m) lubricants or emollients such as petrolatumand lanolin, (n) keratolytic agents (e.g., tazarotene), (O) vitamin D₃derivatives, e.g., calcipotriene or calcipotriol (Dovonex®), (p) PUVA,(q) anthralin (Drithrocreme®), (r) etretinate (Tegison®) andisotretinoin and (s) multiple sclerosis therapeutic agents such asinterferon β-1β (Betaseron®), interferon (β-1α (Avonex®), azathioprine(Imurek®, Imuran®), glatiramer acetate (Capoxone®), a glucocorticoid(e.g., prednisolone) and cyclophosphamide (t) DMARDS such asmethotrexate (u) other compounds such as 5-aminosalicylic acid andprodrugs thereof; hydroxychloroquine; D-penicillamine; antimetabolitessuch as azathioprine, 6-mercaptopurine and methotrexate; DNA synthesisinhibitors such as hydroxyurea and microtubule disrupters such ascolchicine. The weight ratio of the compound of the present invention tothe second active ingredient may be varied and will depend upon theeffective dose of each ingredient. Generally, an effective dose of eachwill be used. Thus, for example, when a compound of the presentinvention is combined with an NSAID the weight ratio of the compound ofthe present invention to the NSAID will generally range from about1000:1 to about 1:1000, preferably about 200:1 to about 1:200.Combinations of a compound of the present invention and other activeingredients will generally also be within the aforementioned range, butin each case, an effective dose of each active ingredient should beused.

VI. Examples

The following examples are offered to illustrate, but not to limit theclaimed invention.

Reagents and solvents used below can be obtained from commercial sourcessuch as Aldrich Chemical Co. (Milwaukee, Wis., USA). ¹H-NMR spectra wererecorded on a Varian Mercury 400 MHz NMR spectrometer. Significant peaksare provided relative to TMS and are tabulated in the order:multiplicity (s, singlet; d, doublet; t, triplet; q, quartet; m,multiplet) and number of protons. Mass spectrometry results are reportedas the ratio of mass over charge, followed by the relative abundance ofeach ion (in parenthesis). In the examples, a single m/e value isreported for the M+H (or, as noted, M−H) ion containing the most commonatomic isotopes. Isotope patterns correspond to the expected formula inall cases. Electrospray ionization (ESI) mass spectrometry analysis wasconducted on a Hewlett-Packard MSD electrospray mass spectrometer usingthe HP1100 HPLC for sample delivery. Normally the analyte was dissolvedin methanol at 0.1 mg/mL and 1 microlitre was infused with the deliverysolvent into the mass spectrometer, which scanned from 100 to 1500daltons. All compounds could be analyzed in the positive ESI mode, usingacetonitrile/water with 1% formic acid as the delivery solvent. Thecompounds provided below could also be analyzed in the negative ESImode, using 2 mM NH₄OAc in acetonitrile/water as delivery system.

Compounds within the scope of this invention can be synthesized asdescribed below, using a variety of reactions known to the skilledartisan. A sample of useful routes to the azaindazole derivatives andcertain compounds of the invention are provided below or elsewherewithin the present application. In the descriptions of the synthesesthat follow, some of the arylpiperazine and heteroaromatic subunitprecursors were obtained from commercial sources. These commercialsources include Aldrich Chemical Co., Acros Organics, Ryan ScientificIncorporated, Oakwood Products Incorporated, Lancaster Chemicals, SigmaChemical Co., Lancaster Chemical Co., TCI-America, Alfa Aesar, DavosChemicals, and GFS Chemicals. Certain relevant arylpiperazine compoundscan be commercially obtained. Others could be prepared as described inU.S. patent application Ser. No. 11/008,774, the contents of which ishereby incorporated in its entirety for all purposes. Also, standardchemistries have been employed to link the arylpiperazine andheteroaromatic subunits (whether commercially obtained or prepared bythe methods below) using a suitably optimized linker, such as the acetylunit described in the body of this invention.

One skilled in the art will also recognize that alternative methods maybe employed to synthesize the target compounds of this invention, andthat the approaches described within the body of this document are notexhaustive, but do provide broadly applicable and practical routes tocompounds of interest.

Certain molecules claimed in this patent can exist in differentenantiomeric and diastereomeric forms and all such variants of thesecompounds are claimed.

Regioisomerism is a common property in organic chemistry, and isespecially common with regards to certain structural types providedherein. Those skilled in the art will recognize, with respect to thecompounds described herein, that the coupling reactions with theheteroaromatic ring systems can lead to either one of or a mixture ofdetectable regioisomers.

The detailed description of the experimental procedures used tosynthesize key compounds in this text lead to molecules that aredescribed by the physical data identifying them as well as by thestructural depictions associated with them.

Those skilled in the art will also recognize that during standard workup procedures in organic chemistry, acids and bases are frequently used.Salts of the parent compounds are sometimes produced, if they possessthe necessary intrinsic acidity or basicity, during the experimentalprocedures described within this patent.

EXAMPLES

General Method A: HATU Coupling.

The desired amine (1.2 equiv) and carboxylic acid were combined in THF(0.2 M) with Et₃N (1-2 equiv) followed by the addition of HATU (1.3equiv) at room temperature. Upon completion of the reaction, the mixturewas diluted with EtOAc, washed with saturated NaHCO₃ solution and brine,dried over Na2SO4, filtered, and concentrated under reduced pressure.

General Method B: Acid Chloride Formation and Coupling.

The desired carboxylic acid was diluted in CHCl₃ (0.25 M) with 2 dropsof DMF. Oxalyl chloride (1.2 equiv) was added at room temperature,giving gas evolution from the mixture. After 20-30 minutes, the desiredamine (1.2 equiv) was added followed by Et₃N (2.5 equiv) or an equalvolume (to CHCl₃) of saturated NaHCO₃ solution. Upon completion of thereaction, the mixture was diluted with EtOAc, washed with NaHCO₃ andbrine, dried over Na2SO4, filtered, and concentrated under reducedpressure.

Example 1: Synthesis of2-[4-amino-3-(1H-imidazol-2-yl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]ethanone

a) To a 500-mL round-bottomed flask was added 4-fluorophenylhydrazinehydrochloride (18.9 g, 116 mmol), 3-fluoro-2-formylpyridine (14.2 g, 113mmol), K₂CO₃(47.0 g, 340 mmol), and DMF (150 mL). The slurry was heatedto 120° C. in an oil bath for 41 hours at which point the reactionmixture was cooled and poured into 1.2 L of H₂O. The tan precipitate wascollected via vacuum filtration and washed with 3×H₂O. The solid wasdissolved in CH₂Cl₂ and the resulting solution was washed with brine,dried over Na2SO₄, filtered, and concentrated under reduced pressure togive a brown solid. The solid was triturated with 200 mL of 9:1hexanes:EtOAc and dried to provide1-(4-fluorophenyl)-1H-pyrazolo[4,3-b]pyridine (16.3 g, 68%) as a tanpowder. MS: (ES) m/calculated for C₁₂H₉FN₃ [M+H]⁺ 214.1, found 214.1.

b) The heterocycle obtained above (16.3 g, 77 mmol) was dissolved in 380mL of methanol and treated with 20% Pd(OH)₂/C (10.7 g) in a 2-L Parrbottle. The system was purged and filled with hydrogen (3×) and thenshaken under 56 psi hydrogen. An additional 8.0 g of catalyst was addedafter 1 day. Once the starting material was consumed, the bottle wasflushed with nitrogen and the reaction mixture filtered through a pad ofCelite, washing with several portions of methanol. The filtrate wasconcentrated under reduced pressure to provide1-(4-fluorophenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyridine (13.7g, 82%) as a light tan solid. MS: (ES) m/z calculated for Cl₂H₁₃FN₃[M+H]⁺ 218.1, found 218.1.

c) The compound was prepared from2-(4-amino-3-(1H-imidazol-2-yl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)aceticacid and1-(4-fluorophenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyridine aboveusing General Method A. The product was purified by reverse phase HPLC(C18 column, acetonitrile-H₂O with 0.1° % TFA as eluent) to give thedesired product as a white solid (6.4 mg). ¹H NMR (400 MHz, MeOH-d₄) δ8.35 (s, 1H), 8.15 (s, 1H), 7.54 (m, 2H), 7.27 (m, 4H), 5.70 (s, 2H),4.00 (m, 2H), 2.91 (t, J=6.2 Hz, 2H), 2.18 (m, 2H); MS: (ES) m/zcalculated for C₂₂H₂₀FN₁₀O [M+H]⁺ 459.18, found 459.1.

Synthesis of2-(4-chloro-5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)acetic acid

a) To a solution of 4-chloro-5-methyl-3-(trifluoromethyl)pyrazole (15 g,81.3 mmol) in DMF (82 mL) was added ethyl 2-bromoacetate (13.6 g, 81.3mmol) and potassium carbonate (12.4 g, 89.4 mmol). The mixture washeated at 55° C. for 10 hours with stirring. After cooling to roomtemperature, the reaction mixture was diluted with water (100 mL) andextracted with ethyl acetate (3×150 mL). The combined organic layerswere washed with brine, dried (Na₂SO₄), filtered, and concentrated invacuo. The crude residue was purified by flash chromatography (SiO₂,0-30% ethyl acetate in hexanes) to afford the desired product (18.5 g,68.4 mmol, 84%).

b) To a solution of ethyl2-(4-chloro-5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)acetate (7.5 g,27.7 mmol) in THF (100 mL) and water (50 mL) was added LiOH.H₂O (2.33 g,55.4 mmol). The mixture was stirred at 60° C. for 5 hours. After coolingto room temperature, the mixture was acidified with 1 N HCl (80 mL) andextracted with 1:2 IPA/CHCl₃ (2×150 mL). The combined organic layerswere washed with brine, dried (Na₂SO₄), filtered, and concentrated invacuo to give the desired compound (6 g, 24.7 mmol, 89%). ¹H NMR (400MHz, DMSO-d₆) δ 4.98 (s, 2H), 2.21 (s, 3H); MS: (ES) m/z calculated forC₇H₆Cl F₃ N₂ O₂ [M+H]⁺243.0, found 243.0.

Example 2: Synthesis of2-[4-chloro-5-methyl-3-(trifluoromethyl)pyrazol-1-yl]-1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]ethanone

The compound was prepared from2-(4-chloro-5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)acetic acid and1-(4-fluorophenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyridine usingGeneral Method B. The product mixture was washed with 2×1 M NaHSO₄ and2×1.5 M KOH and the organics were eluted through a silica plug. Theresulting slurry was concentrated to a white solid that was trituratedwith 1:1 hexanes:EtOAc to give the title compound as a crystalline whitesolid (75.9 mg). ¹H NMR (400 MHz, CDCl₃, mixture of rotamers) δ 8.34 (s,0.7H), 7.57 (s, 0.3H), 7.44 (m, 2H), 7.15 (m, 2H), 5.24 (s, 0.4H), 5.13(s, 1.6H), 3.92 (m, 0.5H), 3.84 (m, 1.5H), 2.87 (m, 2H), 2.33 (s, 2.3H),2.32 (s, 0.7H), 2.12 (m, 1.5H), 2.05 (m, 0.5H); MS: (ES) m/z calculatedfor C₁₉H₁₇ClF₄N₅O [M+H]⁺ 442.1, found 442.0.

Example 3: Synthesis of2-(4-chlorophenyl)-1-[1l-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]ethanone

The compound was from prepared from 4-chlorophenylacetic acid and1-(4-fluorophenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyridine usingGeneral Method A. The product mixture was crystallized from 4:1CH₃CN:H₂O to give the title compound as an off-white crystalline solid(37.6 mg). ¹H NMR (400 MHz, MeOD-d₄, mixture of rotamers) δ 8.18 (s,0.9H), 7.84 (s, 0.1H), 7.56 (m, 2H), 7.25-7.37 (m, 6H), 4.02 (s, 0.2H),3.92 (s, 1.8H), 2.81 (t, J=6.2 Hz, 2H), 2.49 (m, 2H), 1.90 (m, 2H); MS:(ES) m/z calculated for C₂₀H₁₈ClFN₃O [M+H]⁺ 370.1, found 370.1.

Example 4: Synthesis of1-[1-(4-chloro-3-methoxy-phenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-[4-chloro-5-methyl-3-(trifluoromethyl)pyrazol-1-yl]ethanoneand1-(2-(4-chloro-3-methoxyphenyl)-6,7-dihydro-2H-pyrazolo[4,3-b]pyridin-4(5H)-yl)-2-(4-chloro-5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)ethanone

a) 2-(4-Chloro-5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)acetic acid(255 mg, 1.05 mmol) was diluted in 5 mL of CHCl₃ and 2 drops of DMF.Oxalyl chloride (105 μL, 1.2 mmol) was added slowly giving gasevolution. After 30 minutes, 160 mg (1 mmol) of4,5,6,7-tetrahydro-2H-pyrazolo[4,3-b]pyridine*HCl was added followed by0.75 mL of 3 M K₂CO₃. After 30 minutes, 1 mL of 3 M KOH and 1 mL of MeOHwere added to the reaction slurry and the mixture was stirred for 90minutes. The reaction slurry was washed with brine and the organic layerwas dried on MgSO4, filtered, and concentrated to give the amide as atan solid (184 mg, 53%) that was used without further purification. MS:(ES) m/z calculated for C₁₃H₁₄ClF₃N₅O [M+H]⁺ 348.1, found 348.0.

b) The amide formed above (104 mg, 0.3 mmol) was combined with5-bromo-2-chloroanisole (199 mg, 0.9 mmol), 0.3 mL of dioxane, Cu(I)I(11.4 mg, 0.06 mmol, 20%),(±)-trans-N,N′-dimethylcyclohexane-1,2-diamine (21.3 mg, 0.15 mmol,50%), and K₂CO₃ (82.9 mg, 0.6 mmol). The slurry was heated to 120° C.until reaction completion. The product was purified by reverse phaseHPLC (C18 column, acetonitrile-H₂O with 0.1% TFA as eluent) to providetwo regioisomers. First eluting, Compound A:1-[1-(4-chloro-3-methoxy-phenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-[4-chloro-5-methyl-3-(trifluoromethyl)pyrazol-1-yl]ethanonewas isolated as a white foam (8.1 mg). ¹H NMR (400 MHz, CDCl₃, mixtureof rotamers) δ 8.37 (s, 0.7H), 7.60 (s, 0.3H), 7.41 (d, J=8.4 Hz, 1H),7.19 (d, J=2.6 Hz, 1H), 6.92 (dd, J=2.2, 8.4 Hz, 1H), 5.24 (s, 0.5H),5.14 (s, 1.5H), 3.97 (s, 0.6H), 3.95 (s, 2.4H), 3.85 (m, 2H), 2.92 (m,2H), 2.34 (s, 2.6H), 2.33 (s, 0.4H), 2.13 (m, 1.4H), 2.05 (m, 0.6H); MS:(ES) m/z calculated for C₂₀H₁₉F₃N₅O₂ [M+H]⁺ 488.1, found 488.1. Secondeluting, Compound B:1-(2-(4-chloro-3-methoxyphenyl)-6,7-dihydro-2H-pyrazolo[4,3-h]pyridin-4(5H)-yl)-2-(4-chloro-5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)ethanonewas isolated as a white solid (26.8 mg). ¹H NMR (400 MHz, CDCl₃, mixtureof rotamers) δ 8.57 (s, 1H), 7.35 (d, J=8.4 Hz, 1H), 7.26 (d, J=2.2 Hz,1H), 7.11 (dd, J=2.2, 8.4 Hz, 1H), 5.22 (s, 0.2H), 5.16 (s, 1.8H), 3.95(s, 2.5H), 3.86 (s, 0.5H) 3.84 (m, 2H), 2.93 (t, J=6.6 Hz, 2H), 2.32 (s,3H), 2.19 (m, 2H); MS: (ES) m/z calculated for C₂₀H₁₉F₃N₅O₂ [M+H]⁺488.1, found 488.0.

Example 5a: Synthesis of1-[I-(4-chlorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-[4-chloro-5-methyl-3-(trifluoromethyl)pyrazol-1-yl]ethanoneand1-(2-(4-chlorophenyl)-6,7-dihydro-2H-pyrazolo[4,3-b]pyridin-4(5H)-yl)-2-(4-chloro-5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)ethanone

a) 4,5,6,7-Tetrahydro-2H-pyrazolo[4,3-b]pyridine-HCl (319 mg, 2 mmol)was combined with Boc₂O (480 mg, 2.2 mmol) and K₂CO₃ (1.3 mL, 3 Maqueous, 2 equiv) in CH₂Cl₂ (10 mL). After 15 hours, the starting aminewas consumed (LCMS). The reaction slurry was washed with brine and theorganic layer was dried on MgSO₄, filtered, and concentrated to give aviscous yellow oil (446 mg, 96%) that was used without furtherpurification. MS: (ES) m/z calculated for C₁H₁₈N₃O₂ [M+H]⁺ 224.1, found224.2.

b) The Boc-amine formed above (112 mg, 0.5 mmol) was combined with4-chloroiodobenzene (239 mg, 1 mmol), 6 mL of dioxane, Cu(I)I (12 mg,0.06 mmol, 10%), (±)-trans-N,N′-dimethylcyclohexane-1,2-diamine (18.5mg, 0.13 mmol, 25%), and K₂CO₃ (138 mg, 1 mmol). The slurry was heatedto 110° C. overnight. Two regioisomers are obtained in roughly equalquantities. The reaction slurry was diluted with 20 mL of EtOAc andwashed with brine and 2×1 M NaHSO₄. The organic layer was dried overMgSO₄, filtered, and concentrated to an orange oil that was used withoutfurther purification.

c) The regioisomer mixture obtained above (66 mg, approx 0.25 mmoltotal) was combined with2-(4-chloro-5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)acetic acid (67mg, 0.275 mmol), HATU (114 mg, 0.3 mmol), and Et₃N (52 μL, 0.38 mmol) inTHF (1 mL). After 90 minutes the reaction slurry was diluted with EtOAc,washed with H₂O, and concentrated. The residue was purified by reversephase HPLC (C18 column, acetonitrile-H₂O with 0.1% TFA as eluent) toprovide two regioisomers. First eluting, Compound A:1-[1-(4-chlorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-[4-chloro-5-methyl-3-(trifluoromethyl)pyrazol-1-yl]ethanonewas isolated as a white solid (1.7 mg). ¹H NMR (400 MHz, CDCl₃, mixtureof rotamers) δ 8.36 (s, 0.8H), 7.59 (s, 0.2H), 7.43 (s, 4H), 5.24 (s,0.4H), 5.13 (s, 1.6H), 3.92 (m, 0.4H), 3.85 (m, 1.6H), 2.90 (m, 1.8H),2.77 (m, 0.2H), 2.34 (s, 2.4H), 2.32 (s, 0.6H), 2.13 (m, 1.5H), 2.05 (m,0.5H); MS: (ES) m/z calculated for C₁₉H₁₇Cl₂F₃N₅O [M+H]⁺ 458.1, found458.0 Second eluting, Compound B:1-(2-(4-chlorophenyl)-6,7-dihydro-2H-pyrazolo[4,3-b]pyridin-4(5H)-yl)-2-(4-chloro-5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)ethanonewas isolated as a white solid (31.0 mg). ¹H NMR (400 MHz, CDCl₃) δ 8.56(s, 1H), 7.56 (m, 2H), 7.36 (m, 2H), 5.16 (s, 2H), 3.84 (s, 2H), 2.93(t, J=6.6 Hz, 2H), 2.32 (s, 3H), 2.19 (m, 2H), MS: (ES) m/z calculatedfor C₁₉H₁₇Cl₂F₃N₅O [M+H]⁺ 458.1, found 458.0.

Example 5b: Synthesis of1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-pyrazolo[3,4-b]pyridin-1-yl-ethanone

a) To a flask containing1-(4-fluorophenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyridine (0.869g, 4 mmol) and 3 M K₂CO₃(aq) (2.6 mL, 8 mmol) in 10 mL of CH₂Cl₂ wasadded chloroacetyl chloride (0.35 mL, 4.4 mmol) dropwise. After 20minutes, the slurry was diluted with 5 mL of H₂O and the layers wereseparated. The organic layer was washed with brine and 2×1 M NaHSO₄,dried on MgSO₄, filtered, and concentrated to a tan foamy solid (1.05 g,89%). MS: (ES) m/z calculated for C₁₄H₁₄ClFN₃O [M+H]⁺ 294.1, found294.1.

b) The α-chloroacetamide obtained above (147 mg, 0.5 mmol) was dilutedin 1.5 mL of THF and treated with K₂CO₃ (138 mg, 1 mmol) and3-iodo-1H-pyrazolo[3,4-b]pyridine (130 mg, 0.53 mmol). The slurry washeated to 75° C. Upon completion, 4 mL of methanol and 50 mg of 10% Pd/Cwere added to the vessel. The reaction mixture was then shaken on a Parrapparatus under 50 psi of H₂ overnight. The reaction mixture wasfiltered through Celite washing with methanol and concentrated. Theresidue was purified by reverse phase HPLC (C18 column, acetonitrile-H₂Owith 0.1% TFA as eluent) to provide 30.9 mg of product as a whitecrystalline solid. ¹H NMR (400 MHz, CDCl₃, mixture of rotamers) δ 8.54(dd, J=1.5, 4.4 Hz, 1H), 8.35 (s, 0.8H), 8.14 (s, 1H), 8.11 (m, 1H),7.75 (s, 0.2H), 7.43-7.51 (m, 2H), 7.12-7.21 (m, 3H), 5.70 (s, 0.4H),5.59 (s, 1.6H), 3.91 (m, 2H), 2.87 (m, 2H), 2.15 (m, 1.6H), 2.05 (m,0.4H); MS: (ES) m/z calculated for C₂₀H₁₈FN₆O [M+H]⁺ 377.1, found 377.1.

Example 6: Synthesis ofl-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-pyrazolo[3,4-b]pyridin-1-yl-ethanone

a) A septum-capped vial with stirring bar was rinsed with dry THF andcharged with ethyl 4-chloro-5-methyl-1H-pyrazole-3-carboxylate (94 mg,0.5 mmol) and 1 mL of dry THF. Under N₂, the reaction vessel was cooledin an ice bath before dropwise addition of 520 μL of MeMgBr (1.55 mmol,3 M in Et₂O). After 40 minutes, an additional 150 μL of MeMgBr (0.45mmol) was added. Upon completion, the reaction mixture was treated with3 mL of 1 M NaHSO₄ and extracted with 3×EtOAc. The organic layers werecombined, dried on MgSO₄, filtered, and concentrated to provide ayellow-tan solid (69 mg, 79%) that was used without furtherpurification. MS: (ES) m/z calculated for C₇H₁₂ClN₂O [M+H]⁺ 175.1, found175.1.

b) The carbinol obtained above (69 mg, 0.4 mmol) was diluted in 1.2 mLof THF and 500 μL of DMF and treated with K₂CO₃ (111 mg, 0.8 mmol) and2-chloro-1-(1-(4-fluorophenyl)-6,7-dihydro-1H-pyrazolo[4,3-b]pyridin-4(5H)-yl)ethanone(118 mg, 0.4 mmol). The slurry was heated to 75° C. The reaction slurrywas diluted with EtOAc and washed with brine, dried on MgSO₄, filtered,and concentrated. The residue was purified by reverse phase HPLC (C18column, acetonitrile-H₂O with 0.1% TFA as eluent) to provide 69 mg ofthe desired product (43%) as a white solid. ¹H NMR (400 MHz, CDCl₃,mixture of rotamers) δ 8.37 (s, 0.8H), 7.63 (s, 0.2H), 7.42-7.46 (m,2H), 7.13-7.19 (m, 2H), 5.14 (s, 0.4H), 5.03 (s, 1.6H), 3.90 (m, 0.4H),3.87 (m, 1.6H), 2.86 (m, 2H), 2.28 (s, 2.4H), 2.25 (s, 0.6H), 2.05-2.13(m, 2H), 1.62 (s, 6H); MS: (ES) m: calculated for C₂₁H₂₄ClFN₅O₂[M+H]⁺432.2, found 432.1.

Example 7: Synthesis of2-(4-chloro-5-methyl-3-pyrimidin-2-yl-pyrazol-1-yl)-1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]ethanonetrifluoroacetic acid salt and2-(4-chloro-3-methyl-5-pyrimidin-2-yl-pyrazol-1-yl)-1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]ethanonetrifluoroacetic acid salt

2-Chloro-1-(1-(4-fluorophenyl)-6,7-dihydro-1H-pyrazolo[4,3-b]pyridin-4(5H)-yl)ethanone(118 mg, 0.4 mmol) was diluted in 1.2 mL of 2:1 THF:DMF and treated withK₂CO₃ (111 mg, 0.8 mmol) and2-(4-chloro-5-methyl-1H-pyrazol-3-yl)pyrimidine (78 mg, 0.4 mmol). Theslurry was heated to 75° C. and stirred overnight. Two isomers with thesame MW were present as assessed by LCMS. The reaction slurry wasdiluted with EtOAc and washed with brine, dried on MgSO₄, filtered, andconcentrated. The residue was purified by reverse phase HPLC (C18column, acetonitrile-H₂O with 0.1% TFA as eluent) to provide tworegio-isomers.

First Eluting Isomer:

2-(4-Chloro-5-methyl-3-pyrimidin-2-yl-pyrazol-1-yl)-1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]ethanonetrifluoroacetic acid salt: Obtained 25 mg of white needle-shapedcrystals. ¹H NMR (400 MHz, DMSO-d₆) δ 8.86 (d, J=5.1 Hz, 2H), 8.13 (s,1H), 7.59 (m, 2H), 7.44 (t, 0.1=4.8 Hz, 1H), 7.34 (m, 2H), 5.50 (s, 2H),3.84 (m, 2H), 2.87 (t, J=6.2 Hz, 2H), 2.24 (s, 3H), 2.02 (m, 2H): MS:(ES) m/z calculated for C₂₂H₂₀ClFN₇O [M+H]⁺ 452.1, found 452.1. Secondeluting isomer:2-(4-Chloro-3-methyl-5-pyrimidin-2-yl-pyrazol-1-yl)-1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]ethanonetrifluoroacetic acid salt: Obtained 8.8 mg of pale orange solid. ¹H NMR(400 MHz, DMSO-d6, mixture of rotamers) δ 8.87 (d, J=5.1, 1.6H), 8.81(d, l=5.1 Hz, 0.4H), 7.96 (s, 1 H), 7.66 (m, 0.4H), 7.56 (m, 1.6H), 7.42(t, J=4.8, 1H), 7.33 (m, 2H), 5.72 (s, 2H), 3.84 (m, 2H), 2.85 (t, J=6.2Hz, 2H), 2.23 (s, 3H), 1.99 (m, 2H); MS: (ES) m/z calculated forC₂H₂₀ClFN₇O [M+H]⁺ 452.1, found 452.1.

Example 8: Synthesis of1-[i-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-[5-methyl-3-(trifluoromethyl)pyrazol-1-yl]ethanone

2-Chloro-1-(1-(4-fluorophenyl)-6,7-dihydro-1H-pyrazolo[4,3-b]pyridin-4(5)-yl)ethanone(118 mg, 0.4 mmol) was diluted in 1.2 mL of 2:1 THF:DMF and treated withK₂CO₃ (111 mg, 0.8 mmol) and 5-methyl-3-(trifluoromethyl)-1H-pyrazole(60 mg, 0.4 mmol). The slurry was heated to 75° C. for 2 hours. Thereaction slurry was purified by normal phase flash chromatography (24 gcolumn, eluting with 10-80% EtOAc in hexanes) to provide 109 mg (67%) ofthe title compound as a white solid. ¹H NMR (400 MHz, CDCl₃) δ 8.36 (s,0.8H), 7.59 (s, 0.2H), 7.42-7.47 (m, 2H), 7.12-7.25 (m, 2H), 5.25 (s,0.4H), 5.14 (s, 1.6H), 3.92 (m, 0.4H), 3.86 (m, 1.6H), 2.86 (t, 0.1=6.2Hz, 2H), 2.37 (s, 2.4H), 2.36 (m, 0.6H), 2.11 (m, 1.6H), 2.04 (m, 0.4H);MS: (ES) m/z calculated for C₁₉H₁₈F₄N₅O [M+H]⁺ 408.1, found 408.1.

Example 9: Synthesis of2-[4-chloro-5-methyl-3-(trifluoromethyl)pyrazol-1-yl]-1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]propan-1-one

a) Ethyl bromopropionate (1.448 g, 8 mmol),4-chloro-5-methyl-3-(trifluoromethyl)-1H-pyrazole (1.476 g, 8 mmol), andK₂CO₃ (2.211 g, 16 mmol) were combined in 24 mL of 2:1 THF:DMF at 75° C.for 3 hours. The volatiles were evaporated and the residue slurried in20 mL of 4:1 hexanes:EtOAc and washed with 3×H₂O and 2× brine. Theorganic layer was dried and purified by flash chromatography (SiO₂, 80 gcolumn, eluting with 5-80% EtOAc in hexanes) to provide 1.859 g (82%) ofthe title compound as a colorless oil. ¹H NMR (400 MHz, CDCl₃) δ 4.95(q, J=7.3 Hz, 1H), 4.21 (q, J=7.3 Hz, 2H), 2.27 (s, 3H), 1.83 (d, J=7.3Hz, 3H), 1.81 (t, J=7.3 Hz, 1H); MS: (ES) m/z calculated forC₁₀H₁₃ClF₃N₂O₂ [M+H]⁺ 285.1, found 285.0.

b) The ester obtained above was slurried in 13 mL of THF and 6.5 mL of 2M LiOH. Upon completion of the reaction, the volatiles were evaporatedand 9 mL of 2 M HCl was added to precipitate the product. Theprecipitate was washed with water (3×5 mL) and dried under vacuum toconstant weight. The title compound was obtained as a white solid (1.53g, 82%). MS: (ES) m/z calculated for C₈H₉ClF₃N₂O₂[M+H]⁺ 257.0, found257.0.

c) The title compound was prepared from2-(4-chloro-5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)propanoic acidand 1-(4-fluorophenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyridineusing General Method B. The product mixture was washed with 2×1 MNaHSO₄. The reaction slurry was purified by flash chromatography (SiO₂,24 g column, eluting with 5-50% EtOAc in hexanes) to provide 137 mg(75%) of the title compound as a white foam. ¹H NMR (400 MHz, CDCl₃) δ8.41 (s, 1H), 7.43 (m, 2H), 7.15 (m, 2H), 5.52 (q, J=7.0 Hz, 1H), 3.75(m, 1H), 3.41 (m, 1H), 2.81 (m, 2H), 2.28 (s, 3H), 1.85-1.99 (m, 2H),1.81 (d, J=7 Hz, 1H); MS: (ES) m/z calculated for C₂₀H₁₉ClF₄N₅O [M+H]⁺456.1, found 456.1.

Example 10: Synthesis of2-(4-chlorophenyl)-1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]ethanone

The compound was prepared from 3-trifluoromethylphenylacetic acid and1-(4-fluorophenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-h]pyridine usingGeneral Method B. The product mixture was washed with 2×1 M NaHSO₄. Thereaction slurry was purified by flash chromatography (SiO₂, 24 g column,eluting with 5-50% EtOAc in hexanes) to provide 83 mg (69%) of the titlecompound as a white solid. ¹H NMR (400 MHz, CDCl₃, mixture of rotamers)δ 8.45 (s, 1H), 7.42-7.55 (m, 6H), 7.14 (m, 2H), 4.08 (s, 0.3H), 3.95(s, 1.7H), 3.77 (m, 2H), 2.83 (t, J=6.6 Hz, 2H), 2.01 (m, 2H); MS: (ES)m/z calculated for C₂₁H₁₈F₄N₃O [M+H]⁺ 404.1, found 404.1.

Example 11: Synthesis of2-[4-chloro-5-methyl-3-(trifluoromethyl)pyrazol-1-yl]-1-[1-(4-fluorophenyl)-3-methyl-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]ethanone

a) To a 100-mL roundbottomed flask was added 4-fluorophenylhydrazinehydrochloride (3.33 g, 20.5 mmol), K₂CO₃ (8.29 g, 60 mmol), and DMF (27mL). To this slurry was added 2.78 g (20 mmol) of1-(3-fluoropyridin-2-yl)ethanone, giving a yellow suspension. Themixture was heated to 120° C. for 24 hours at which point the startingmaterials were consumed. The slurry was cooled to room temperature and70 mL of H₂O was added to precipitate the product. The solid wascollected on a medium frit and washed with 3×H₂O. The solid was dried at60° C. to provide the product as a tan solid (3.82 g, 84%). MS: (ES) m/zcalculated for C₁₃H₁₁FN₃ [M+H]⁺ 228.1, found 228.1.

b) Methanol (80 mL), 20% Pd(OH)₂/C (472 mg, approx 0.33 mmol), and1-(4-fluorophenyl)-3-methyl-1H-pyrazolo[4,3-b]pyridine (3.82 g, 16.8mmol) were combined in a Parr flask and purged with H₂. The flask wascharged to 55 psi H₂ and then the slurry was shaken overnight. Themixture was filtered through Celite, washing with several portions ofmethanol. The filtrate was then concentrated to give 3.73 g (96% crudeyield) of an orange oil that solidified on standing. MS: (ES) m/zcalculated for C₁₃H₁₅FN₃ [M+H]⁺ 232.1, found 232.1.

c) The title compound was prepared from2-(4-chloro-5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)acetic acid and1-(4-fluorophenyl)-3-methyl-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyridineusing General Method B. The product mixture was washed with 1 M NaHSO₄.The reaction slurry was purified by flash chromatography (SiO₂, 24 gcolumn, eluting with 5-50% EtOAc in hexanes) to provide 75 mg (41%) ofthe title compound as a white solid. ¹H NMR (400 MHz, CDCl₃, mixture ofrotamers) δ 7.47 (m, 0.8H), 7.40 (m, 1.2H), 7.14 (m, 2H), 5.11 (s,1.2H), 5.01 (s, 0.8H), 3.81 (m, 2H), 2.85 (m, 2H), 2.29-2.40 (m, 6H),2.07 (m, 2H); MS: (ES) m/z calculated for C₂₀H₁₉ClF₄N₅O [M+H]⁺ 456.1,found 456.1.

Example 12: Synthesis of1-(1-(4-fluorophenyl)-6,7-dihydro-1H-pyrazolo[4,3-b]pyridin-4(5H)-yl)-2-(2-(trifluoromethyl)phenyl)ethanone

The compound was prepared from 2-trifluoromethylphenylacetic acid and1-(4-fluorophenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyridine usingGeneral Method B. The product mixture was washed with 1 M NaHSO₄. Thereaction slurry was purified by flash chromatography (SiO₂, 24 g column,eluting with 5-50%0 EtOAc in hexanes) to provide 72 mg (59%) of thetitle compound as a white solid. ¹H NMR (400 MHz, CDCl₃, mixture ofrotamers) δ 8.46 (s, 0.8H), 7.70 (m, 1H), 7.55 (m, 1.2H), 7.41-7.50 (m,4H), 7.15 (m, 2H), 4.19 (s, 0.3H), 4.06 (s, 1.7H), 3.95 (m, 0.3H), 3.76(m, 1.7H), 2.82-2.88 (m, 2H), 2.03 (m, 2H); MS: (ES) m/z calculated forC₂₁H₁₈F₄N₃O [M+H]⁺ 404.1, found 404.1.

Example 13: Synthesis of1-(1-(4-fluorophenyl)-6,7-dihydro-1H-pyrazolo[4,3-b]pyridin-4(5H)-yl)-2-(4-(trifluoromethyl)phenyl)ethanone

The compound was prepared from 4-trifluoromethylphenylacetic acid and1-(4-fluorophenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyridine usingGeneral Method B. The product mixture was washed with 1 M NaHSO₄. Thereaction slurry was purified by flash chromatography (SiO₂, 24 g column,eluting with 5-50% EtOAc in hexanes) to provide 77 mg (64%) of the titlecompound as a white solid. ¹H NMR (400 MHz, CDCl₃, mixture of rotamers)δ 8.45 (s, 0.8H), 7.62 (d, J=7.8 Hz, 2H), 7.55 (s, 0.2H), 7.40-7.49 (m,4H), 7.15 (m, 2H), 4.08 (s, 0.3H), 3.96 (m, 2H), 3.75 (m, 1.7H),2.80-2.86 (m, 2H), 2.00 (m, 2H); MS: (ES) m/z calculated for C₂₁H₁₈F₄N₃O[M+H]⁺ 404.1, found 404.1.

Example 14: Synthesis of2-(4-chloro-3-(trifluoromethyl)phenyl)-1-(1-(4-fluorophenyl)-6,7-dihydro-1H-pyrazolo[4,3-b]pyridin-4(5H)-yl)ethanone

The compound was prepared from 4-chloro-3-trifluoromethylphenylaceticacid and1-(4-fluorophenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyridine usingGeneral Method B. The product mixture was washed with brine and 2×1 MNaHSO₄. The reaction slurry was purified by flash chromatography (SiO₂,24 g column, eluting with 5-50% EtOAc in hexanes) to provide 54 mg (41%)of the title compound as a white solid. ¹H NMR (400 MHz, CDCl₃, mixtureof rotamers) δ 8.44 (s, 0.8H), 7.59 (m, 1H), 7.57 (s, 0.2H), 7.41-7.51(m, 4H), 7.17 (m, 2H), 4.03 (s, 0.3H), 3.95 (m, 0.3H), 3.91 (s, 1.7H),3.78 (m, 1.7H), 2.82-2.85 (m, 2H), 2.05 (m, 2H); MS: (ES) m/z calculatedfor C₂₁H₁₇ClF₄N₃O [M+H]⁺ 438.1, found 438.1.

Example 15: Synthesis of2-(3-chloro-4-(trifluoromethyl)phenyl)-1-(1-(4-fluorophenyl)-6,7-dihydro-1H-pyrazolo[4,3-b]pyridin-4(5H)-yl)ethanone

The compound was from prepared from3-chloro-4-trifluoromethylphenylacetic acid and1-(4-fluorophenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyridine usingGeneral Method B. The product mixture was washed with brine and 2×1 MNaHSO₄. The reaction slurry was purified by flash chromatography (SiO₂,24 g column, eluting with 5-60% EtOAc in hexanes) to provide 68 mg (52%)of the title compound as a white solid. ¹H NMR (400 MHz, CDCl₃, mixtureof rotamers) δ 8.45 (s, 0.8H), 7.67 (m, 1H), 7.55 (s, 0.2H), 7.44-7.49(m, 3H), 7.30 (m, 1H), 7.14-7.21 (m, 2H), 4.04 (s, 0.4H), 3.94 (m,0.4H), 3.92 (s, 1.6H), 3.77 (m, 1.6H), 2.82-2.87 (m, 2H), 2.04 (m, 2H);MS: (ES) m/z calculated for C₂₁H₁₇ClF₄N₃O [M+H]⁺ 438.1, found 438.1.

Example 16: Synthesis of2-[4-chloro-5-methyl-3-(trifluoromethyl)pyrazol-1-yl]-1-[1-(4-fluorophenyl)-3-methyl-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]ethanone

a) 2,6-Lutidine-α-2,3-diol (4.17 g, 30 mmol) was dissolved in 60 mL ofCHCl₃ and treated with MnO₂ (15.65 g, 180 mmol). The slurry was heatedto 62° C. for 90 minutes at which point the reaction was complete. Thereaction mixture was filtered through a thin layer of Celite and washedwith several portions of CHCl₃. The filtrate was concentrated to providethe product as a tan crystalline solid (3.24 g, 79%). MS: (ES) m/zcalculated for C₇H₈NO₂ [M+H]⁺ 138.0, found 138.0.

b) The aldehyde obtained above was dissolved in CH₂Cl₂ (20 mL) and Et₃N(2.3 mL, 16.5 mmol). Trifluoromethanesulfonic anhydride (Tf₂O, 2.78 mL,16.5 mmol) was added dropwise over 5 minutes. Additional portions ofEt₃N (500 μL) and Tf₂O (200 μL) were added after 30 minutes. After 75minutes, the reaction mixture was washed with 2×H₂O and 2×saturatedNaHCO₃. The organic layer was dried on MgSO₄, filtered, and concentratedto provide 3.02 g (75%) of orange oil that was used directly in the nextstep.

c) The triflate above (1.08 g, 4 mmol) was dissolved in 5 mL of DMF in aseptum-capped vial. Potassium carbonate (1.66 g, 12 mmol) and4-fluorophenylhydrazine hydrochloride (678 mg, 4.17 mmol) were added andthe slurry was stirred at room temperature, becoming a deep orange-red.Within 3 hours, LCMS indicated that the starting material was consumedproviding the hydrazine intermediate (both with and without triflate).To this reaction mixture was added 400 mg (0.55 mmol) of PdCl₂(dppf) and5 mL of PhCH₃. The flask was placed in a 100° C. heating blockovernight. The reaction mixture was poured into 100 mL of H₂O and 80 mLof EtOAc. The mixture was filtered through a plug of Celite and thelayers separated. The organic layer was concentrated and the residue waspurified by normal phase flash chromatography (40 g column, eluting with15-70% EtOAc in hexanes) to provide 160 mg (18%) of the title compoundas an orange oily solid. MS: (ES) m/z calculated for Cl₃H₁₁FN₃ [M+H]⁺228.1, found 228.1.

d) Methanol (3.5 mL), 20% Pd(OH)₂/C (200 mg, approx 0.14 mmol), and1-(4-fluorophenyl)-5-methyl-1H-pyrazolo[4,3-b]pyridine (160 mg, 0.7mmol) were combined in a Parr flask and purged 2×H₂. The flask wascharged to 55 psi H₂ and then the slurry was shaken for 2 days. Themixture was filtered through Celite washing with several portions ofmethanol. The filtrate was concentrated to 103 mg (64% crude yield) ofan orange oily solid. MS: (ES) m/z calculated for C₁₃H₁₅FN₃ [M+H]⁺232.1, found 232.1.

e) The compound was from prepared from from2-(4-chloro-5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)acetic acid and1-(4-fluorophenyl)-5-methyl-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyridineusing General Method B. The product mixture was washed with 2×1 MNaHSO₄. The reaction slurry was purified by flash chromatography (SiO₂,24 g column, eluting with 5-50% EtOAc in hexanes) to provide 37 mg (37%)of the title compound as a white foam. ¹H NMR (400 MHz, CDCl₃, mixtureof rotamers) δ 8.37 (s, 0.8H), 7.58 (s, 0.2H), 7.49 (m, 2H), 7.17 (m,2H), 5.08-5.31 (m, 2H), 4.51 (m, 1H), 2.78-2.99 (m, 2 H), 2.34 (s,2.3H), 2.31 (s, 0.7H), 1.97-2.09 (m, 2H), 1.36 (d, J=6.7 Hz, 2.4H), 1.15(d, J=6.6 Hz, 0.6H); MS: (ES) m/z calculated for C₂₀H₁₉ClF₄N₅O [M+H]⁺456.1, found 456.1.

General Scheme for Examples 17-20

Example 17: Synthesis of1-(1-(4-fluorophenyl)-6,7-dihydro-1H-pyrazolo[4,3-b]pyridin-4(5H)-yl)-2-(4-(trifluoromethyl)-1H-imidazol-1-yl)ethanonetrifluoroacetic acid salt

a) To a vial was added 4-trifluoromethyl-1H-imidazole (953 mg, 7 mmol),7 mL of ethanol, and sodium ethoxide (524 mg, 7.7 mmol). To this mixturewas slowly added ethyl bromoacetate (1.29 g, 7.7 mmol). After 18 hours,an additional 100 μL of ethyl bromoacetate and approximately 100-200 mgof NaOEt were added. After 90 minutes, the reaction was quenched withNaHCO₃ and extracted with 3×EtOAc. The organic layer was concentratedand the residue was purified by flash chromatography (SiO₂, 80 g column,eluting with 15-90% EtOAc in hexanes) to provide 746 mg (48%) of thetitle compound as a crystalline solid. MS: (ES) m/z calculated forC₈H₁₀F₃N₂O₂ [M+H]⁺ 223.1, found 223.1.

b) The ester obtained above (746 mg, 3.35 mmol) was dissolved in 10 mLof THF and treated with 3.5 mL of 2 M LiOH(aq). The mixture was heatedin a 50° C. bath for 5 hours. The volatiles were evaporated and 1 mL of6 N HCl (aq) and 1 mL of 1 M NaHSO₄ were added. The resultingprecipitate was collected and washed with H₂O, dissolved in EtOAc, andconcentrated to white solid (200 mg). The aqueous washes wereconcentrated to half volume and a second crop was collected (141 mg). ¹HNMR (400 MHz, CD₃OD) δ 7.80 (s, 1H), 7.64 (s, 1H), 4.94 (s, 2H); MS:(ES) m/z calculated for C₆H₆F₃N₂O₂ [M+H]⁺ 195.0, found 195.1 Theregiochemistry of the system was confirmed by nOe between theα-methylene and both imidazole ring protons.

c) The title compound was prepared from2-(4-(trifluoromethyl)-1H-imidazol-1-yl)acetic acid and1-(4-fluorophenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyridine usingGeneral Method B. The product mixture was washed with 2×brine. Thereaction slurry was purified by reverse phase HPLC (C18 column,acetonitrile-H₂O with 0.1% TFA as eluent) to provide 29 mg of the titlecompound as a white solid. ¹H NMR (400 MHz, CD₃OD, mixture of rotamers)δ 8.25 (s, 0.9H), 7.85 (s, 0.1H), 7.82 (s, 1H), 7.65 (s, 1H), 7.55 (m,2H), 7.27 (m, 2H), 5.45 (s, 0.2H), 5.33 (s, 1.8H), 3.90 (m, 0.2H), 3.87(m, 1.8H), 2.88 (t, J=6.6 Hz, 2H), 2.13 (m, 1.8H), 2.03 (m, 0.2H); MS:(ES) m/z calculated for C₁₈H₁₆F₄N₅O [M+H]⁺ 394.1, found 394.1.

Example 18: Synthesis of1-(1-(4-fluorophenyl)-6,7-dihydro-1H-pyrazolo[4,3-b]pyridin-4(5H)-yl)-2-(2-methyl-4-(trifluoromethyl)-1H-imidazol-1-yl)ethanonetrifluoroacetic acid salt

a) To a vial was added 2-methyl-4-(trifluoromethyl)-1H-imidazole (750mg, 5 mmol), 5 mL of THF, 2.5 mL of DMF, and K₂CO₃ (2.07 g, 15 mmol). Tothis mixture was slowly added ethyl bromoacetate (1.00 g, 6 mmol). After2 hours, the slurry was diluted with EtOAc, filtered on Celite, andconcentrated. The residue was purified by flash chromatography (SiO₂, 80g column, eluting with 15-100% EtOAc in hexanes) to provide 476 mg (40%)of the title compound as a colorless oil. ¹H NMR (400 MHz, CDCl₃) δ 7.20(s, 1H), 4.61 (s, 2H), 4.28 (q, J=7.0 Hz, 2H), 1.63 (s, 3H), 1.31 (t,J=7.0 Hz, 3H); MS: (ES) m/z calculated for C₉H₁₂F₃N₂O₂ [M+H]⁺ 237.1,found 237.1. The regiochemistry of the system was confirmed by nOebetween the α-methylene and both the imidazole ring and methyl protons.

b) The ester obtained above (476 mg, 2 mmol) was dissolved in 6 mL ofTHF and treated with 2 mL of 2 M LiOH the resulting mixture was stirredat room temperature overnight. The volatiles were then evaporated and670 μL of 6 N HCl (aq) was added. The resulting precipitate wascollected, washed with H₂O, and dried under vacuum to constant weight toprovide the title compound as a white powder (284 mg, 68%). MS: (ES) m/zcalculated for C₆H₆F₃N₂O₂ [M+H]⁺ 209.0, found 209.0.

c) The title compound was prepared from from2-(2-methyl-4-(trifluoromethyl)-1H-imidazol-1-yl)acetic acid and1-(4-fluorophenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyridine usingGeneral Method B. The product mixture was diluted with 1 mL of CHCl₃ andwashed with H₂O. The organic layer was concentrated and the residue waspurified by reverse phase HPLC (C18 column, acetonitrile-H₂O with 0.1%TFA as eluent) to provide 31 mg of the title compound as a white solid.¹H NMR (400 MHz, CD₃OD, mixture of rotamers) δ 8.24 (s, 0.9H), 7.88 (s,0.1H), 7.52-7.60 (m, 3H), 7.24-7.32 (m, 2H), 5.36 (s, 0.2H), 5.26 (s,1.8H), 3.89 (m, 2H), 2.89 (t, J=6.2 Hz, 2H), 2.38 (s, 2.7H), 2.36 (m,0.3H), 2.14 (m, 2H); MS: (ES) m/z calculated for C₁₉H₁₈F₄N₅O [M+H]⁺408.1, found 408.1.

Example 19: Synthesis of1-(I-(4-fluorophenyl)-6,7-dihydro-H-pyrazolo[4,3-b]pyridin-4(5H)-yl)-2-(3-(trifluoromethyl)-1H-1,2,4-triazol-1-yl)ethanone

a) To a vial was added 3-(trifluoromethyl)-1H-1,2,4-triazole (358 mg,2.6 mmol), 2.6 mL of ethanol, and sodium ethoxide (354 mg, 5.2 mmol). Tothis mixture was slowly added ethyl bromoacetate (320 μL, 2.9 mmol).After 18 hours, the reaction mixture was concentrated and acidified with1 mL of 6 N HCl (aq). The slurry was diluted in EtOAc and H₂O and thelayers were separated. The organic layer was concentrated and theresidue was purified by flash chromatography (SiO₂, 40 g column, elutingwith 2-80/o EtOAc in hexanes) to provide 423 mg (73%) of the titlecompound as a crystalline solid. ¹H NMR (400 MHz, CDCl₃) δ 8.30 (s, 1H),5.05 (s, 2H), 4.29 (q, J=7.3 Hz, 2H), 1.32 (t, J=7.1 Hz, 3H); MS: (ES)m/z calculated for C₈H₁₀F₃N₂O₂ [M+H]⁺ 224.1, found 224.1. Theregiochemistry of the system was confirmed by nOe between theα-methylene and triazole ring proton.

b) The ester obtained above (142 mg, 0.64 mmol) was dissolved in 2 mL ofTHF and treated with 650 μL of 2 M LiOH (aq). The mixture was stirred at50° C. for 75 minutes. The volatiles were evaporated and 500 μL of 6 NHCl (aq) was added. The solution was extracted with 2×EtOAc, dried onMgSO₄, filtered, and concentrated to provide the title compound as acolorless oil (108 mg, 76%). MS: (ES) m/z calculated for C₅H3F₃N₃O₂[M−H]⁻ 194.1, found 194.1.

c) The title compound was prepared from2-(3-(trifluoromethyl)-1H-1,2,4-triazol-1-yl)acetic acid and1-(4-fluorophenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyridine usingGeneral Method B. The product mixture was washed with 2×brine. Thereaction slurry was purified by reverse phase HPLC (C18 column,acetonitrile-H₂O with 0.1% TFA as eluent) to provide 33 mg of the titlecompound as a white solid. ¹H NMR (400 MHz, CD₃OD, mixture of rotamers)δ 8.41 (s, 1H), 8.35 (s, 0.75H), 7.60 (s, 0.25H), 7.43-7.50 (m, 2H),7.15-7.23 (m, 2H), 5.41 (s, 0.5H), 5.30 (s, 1.5H), 3.94 (m, 0.5H), 3.84(m, 1.5H), 2.89 (m, 2H), 2.15 (m, 1.5H), 2.07 (m, 0.5H); MS: (ES) m/zcalculated for C₁₇H₁₅F₄N₆O [M+H]⁺ 395.1, found 395.1.

Example 20: Synthesis of1-(1-(4-fluorophenyl)-6,7-dihydro-1H-pyrazolo[4,3-b]pyridin-4(5H)-yl)-2-(5-methyl-3-(trifluoromethyl)-1H-1,2,4-triazol-1-yl)ethanone

a) To a flask was added 5-methyl-3-(trifluoromethyl)-1H-1,2,4-triazole(2.27 g, 15 mmol), 15 mL of EtOH, and sodium ethoxide (2.04 g, 30 mmol).To this mixture was slowly added ethyl bromoacetate (2.76 g, 16.5 mmol).After 5 hours, the reaction was quenched with 3 mL of 6 N HCl (aq) andadjusted to pH 7 with saturated NaHCO₃. The volume was reduced by halfand the slurry was diluted with 30 mL of EtOAc. The layers wereseparated and the organic layer washed with H₂O. The organic layer wasconcentrated and the residue was purified by flash chromatography (SiO₂,80 g column, eluting with 5-60% EtOAc in hexanes) to provide 2.00 g(56%) of the title compound as a pale yellow oil. ¹H NMR (400 MHz,CDCl₃) 4.91 (s, 2H), 4.28 (q, J=7.0 Hz, 2H), 2.50 (s, 3H), 1.31 (t,J=7.3 Hz, 3H); MS: (ES) m/z calculated for C₈H₁₁F₃N₃O₂[M+H]⁺ 238.1,found 238.1. The regiochemistry of the system was confirmed by nOebetween the α-methylene and both the triazole methyl protons.

b) The ester obtained above (2.00 g, 8.4 mmol) was dissolved in 10 mL ofTHF and treated with 8 mL of 2 M LiOH (aq). The mixture was stirred at50° C. for 2 hours. The volatiles were evaporated and 2.5 mL of 6 N HCl(aq) was added. The slurry was extracted with 20 mL of EtOAc, dried onMgSO₄, filtered, and concentrated to provide the title compound (1.59 g,90%) as yellow oil that solidified on standing. MS: (ES) m/z calculatedfor C₆H5F₃N₃O₂ [M−H]⁻ 208.0, found 208.1.

c) The title compound was prepared from2-(5-methyl-3-(trifluoromethyl)-1H-1,2,4-triazol-1-yl)acetic acid and1-(4-fluorophenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyridine usingGeneral Method B. The product mixture was diluted with 1 mL of CHCl₃ andwashed with H₂O. The organic layer was concentrated and the residue waspurified by reverse phase HPLC (C18 column, acetonitrile-H₂O with 0.1%TFA as eluent) to provide 37 mg of the title compound as a white solid.¹H NMR (400 MHz, CDCl₃, mixture of rotamers) δ 8.34 (s, 0.8H), 7.59 (s,0.2H), 7.44-7.50 (m, 2H), 7.14-7.27 (m, 2H), 5.29 (s, 0.4H), 5.18 (s,1.6H), 3.93 (m, 0.4H), 3.86 (m, 1.6H), 2.88 (m, 2H), 2.56 (s, 2.4H),2.55 (m, 0.6H), 2.14 (m, 1.6H), 2.06 (m, 0.4H); MS: (ES) m/z calculatedfor C₁₈H¹⁷F₄N₆O [M+H]⁺ 409.1, found 409.1.

Example 21: Synthesis of2-(4-chloro-5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)-1-(1-(5-fluoropyridin-2-yl)-6,7-dihydro-1H-pyrazolo[4,3-b]pyridin-4(5H)-yl)ethanone

2-(4-Chloro-5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)-1-(6,7-dihydro-1H-pyrazolo[4,3-b]pyridin-4(5H)-yl)ethanone(104 mg, 0.3 mmol) was combined with 5-fluoro-2-bromopyridine (106 mg,0.6 mmol), 0.3 mL of dioxane, Cu(I)I (11.4 mg, 0.06 mmol, 20%),(±)-trans-N,N′-dimethylcyclohexane-1,2-diamine (21.3 mg, 0.15 mmol,50%), and K₂CO₃ (83 mg, 0.6 mmol). The slurry was heated to 120° C.until complete (approx 3 hours). The reaction mixture was diluted in 4mL of EtOAc and washed with 2×H₂O. The organic layer was concentratedand the residue was purified by reverse phase HPLC (C18 column,acetonitrile-H₂O with 0.1% TFA as eluent) to provide the title compoundas a white solid. ¹H NMR (400 MHz, CDCl₃, mixture of rotamers) δ 8.91(s, 0.8H), 8.33 (d, J=2.7 Hz, 0.2H), 8.28 (d, J=2.9 Hz, 0.8H), 7.87-7.94(m, 1H), 7.68-7.77 (m, 1H), 5.42 (s, 1.6H), 5.41 (s, 0.4H), 3.93 (s,1.6H), 3.88 (s, 0.4H), 2.88 (t, J=6.4 Hz, 2H), 2.29 (s, 0.6H), 2.28 (s,2.4H), 2.18 (m, 2H); MS: (ES) m/z calculated for C₁₈H₁₆ClF₄N₆O [M+H]⁺443.1, found 443.0.

Example 22: Synthesis of ethyl2-(4-chloro-5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)-2-methylpropanoateand ethyl3-(4-chloro-5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)-2-methylpropanoate

To 4-chloro-5-methyl-3-(trifluoromethyl)-1H-pyrazole (1.846 g, 2.22 mL,10 mmol) and cesium carbonate (6.516 g, 20 mmol) slurried in DMF (20 mL)was added ethyl α-bromoisobutyrate (2.926 g, 15 mmol) in one portion.The flask was placed in an oil bath and the system heated to 50° C.After 2 hours, an additional portion of ethyl α-bromoisobutyrate (1 mL)was added and heating continued. Upon completion as determined byconsumption of the pyrazole, the slurry was diluted with EtOAc andfiltered through Celite, washing with an additional 150 mL of EtOAc. Thefiltrate was washed with 3×brine (half saturated), dried on MgSO₄,filtered, and concentrated to a yellow oil. The product was purified byflash chromatography (SiO₂, 150 g column, eluting with 5-60% EtOAc inhexanes) to provide 1.24 g (42%) of the geminal dimethyl compound and434 mg (15%) of the α-methyl compound as colorless oils. Ethyl2-(4-chloro-5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)-2-methylpropanoate.¹H NMR (400 MHz, CDCl₃) δ 4.25 (q, J=7.1 Hz, 2H), 2.19 (s, 3H), 1.81 (s,6H), 1.27 (t, J=7.1 Hz, 3H); MS: (ES) m/z calculated for C₁₁H5ClF₃N₂O₂[M+H]⁺ 299.1, found 299.0. The regiochemistry of the system wasconfirmed by nOe between the α-methyl and pyrazole methyl protons. Ethyl3-(4-chloro-5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)-2-methylpropanoate.¹H NMR (400 MHz, CDCl₃) 4.35 (dd, J=8.2, 13.8 Hz, 1H), 4.11 (q, J=7.1Hz, 2H), 4.02 (dd, J=6.5, 13.8 Hz, 1H), 3.18 (m, 1H), 2.30 (s, 3H), 1.21(t, J=7.0 Hz, 3H), 1.21 (d, J=7.1 Hz, 3H); MS: (ES) m/z calculated forC₁₁H₁₅ClF₃N₂O₂ [M+H]⁺ 299.1, found 299.0. The regiochemistry of thesystem was confirmed by nOe between the pyrazole methyl and neighboringprotons.

Example 23: Synthesis of2-[4-chloro-5-methyl-3-(trifluoromethyl)pyrazol-1-yl]-1-[I-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-methyl-propan-1-one

a) To ethyl2-(4-chloro-5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)-2-methylpropanoate(1.24 g, 4.2 mmol) was added 12 mL of THF and 4.2 mL of 2 M LiOH (aq).The slurry was heated at 50° C. for 80 min and then the volume reducedby half. The solution was acidified with HCl to provide a whiteprecipitate that was washed with 1 M NaHSO₄ and 2×H₂O. Drying provided1.06 g (94%) the title compound. MS: (ES) m/z calculated forC₉H₁₁ClF₃N₂O₂ [M+H]⁺ 271.0, found 271.0.

b) The title compound was from prepared from from2-(4-chloro-5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)-2-methylpropanoicacid and1-(4-fluorophenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyridine usingGeneral Method B. The product mixture was washed with 2×1 M NaHSO₄. Theorganic layer was concentrated and the residue was purified by flashchromatography (SiO₂, 24 g column, eluting with 5-50% EtOAc in hexanes)to provide 80 mg (68%) of the title compound as a white solid. ¹H NMR(400 MHz, CDCl₃) δ 8.44 (s, 1H), 7.44 (m, 2H), 7.16 (m, 2H), 3.08 (m,2H), 2.74 (t, J=6.6 Hz, 2H), 2.21 (s, 3H), 1.91 (s, 6H), 1.76 (ddd,J=5.8, 6.2, 10.5 Hz, 2H); MS: (ES) m/z calculated for C₂₁H₂₁ClF₄N₅O[M+H]⁺ 470.1, found 470.1.

General Scheme for Examples 24 and 25

Example 24: Synthesis of2-(4-chloro-5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)-1-(1-(4-fluorophenyl)-6,7-dihydro-1H-pyrazolo[4,3-b]pyridin-4(5H)-yl)butan-1-one

a) Methyl bromobutyrate (996 mg, 5.5 mmol),4-chloro-5-methyl-3-(trifluoromethyl)-1H-pyrazole (923 mg, 5 mmol), andK₂CO₃ (1.382 g, 10 mmol) were combined in 15 mL of 2:1 THF:DMF at 50° C.overnight. The solids were filtered off washing with EtOAc and thevolatiles were evaporated. The residue was diluted in EtOAc and washedwith 3×H₂O and 2×brine. The organic layer was concentrated and theresidue was purified by flash chromatography (SiO₂, 40 g column, elutingwith 2-50% EtOAc in hexanes) to provide 1.06 g (74%) of the product as acolorless oil. ¹H NMR (400 MHz, CDCl₃) 4.27 (dd, J=6.7, 9.0 Hz, 1H),3.74 (s, 3H), 2.34 (m, 2H), 2.27 (s, 3H), 0.89 (t, J=6.5 Hz, 3H); MS:(ES) m/z calculated for C₁₀H₁₃ClF₃N₂O₂ [M+H]⁺ 285.1, found 285.0. Theregiochemistry of the system was confirmed by nOe between the α-methineproton and pyrazole methyl protons (and those of the ethyl group).

b) The ester obtained above (1.06 g, 3.72 mmol) was slurried in 10 mLTHF and 3.7 mL of 2 M LiOH (aq) at 50° C. After 70 minutes, thevolatiles were evaporated and 1.25 mL of 6 M HCl (aq) was added. Theslurry was extracted with 2×CH₂Cl₂ and the organic layer dried on MgSO₄,filtered, and concentrated to a pale yellow solid (0.844 mg, 84%). MS:(ES) m/z calculated for C₉H₁₁ClF₃N₂O₂ [M+H]⁺ 270.0, found 270.0.

c) The title compound was prepared from2-(4-chloro-5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)butanoic acidand 1-(4-fluorophenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyridineusing General Method B. The product mixture was washed with 2×1 MNaHSO₄. The reaction slurry was purified by flash chromatography (SiO₂,24 g column, eluting with 5-50% EtOAc in hexanes) to provide 81 mg (69%)of the title compound as a white solid. ¹H NMR (400 MHz, CDCl₃) δ 8.43(s, 1H), 7.44 (m, 2H), 7.16 (m, 2H), 5.30 (dd, J=6.7, 8.6 Hz, 1H), 3.81(ddd, J=3.2, 8.6, 11.5 Hz, 1H), 3.50 (ddd, J=3.1, 7.4, 10.2 Hz, 1H),2.79 (dt, J=2.8, 6.3 Hz, 2H), 2.33 (m, 1H), 2.30 (s, 3H), 2.22 (m, 1H),1.96 (m, 1H), 1.86 (m, 1H), 0.98 (t, J=7.0 Hz, 3H); MS: (ES) m/zcalculated for C₂₀H₁₉ClF₄N₅O [M+H]⁺ 470.1, found 470.1.

Example 25: Synthesis of2-[4-chloro-5-methyl-3-(trifluoromethyl)pyrazol-1-yl]-1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-3-methyl-butan-1-one

a) Ethyl 2-bromo-3-methylbutanoate (836 mg, 4 mmol),4-chloro-5-methyl-3-(trifluoromethyl)-1H-pyrazole (738 mg, 4 mmol), andK₂CO₃(1.106 g, 8 mmol) were combined in 12 mL of 2:1 THF:DMF at 50° C.overnight. An additional 400 μL of alkylator was added and stirringcontinued overnight. The solids were filtered off washing with EtOAc andthe volatiles were evaporated. The residue was diluted in EtOAc andwashed with 2×brine (half-saturated). The organic layer was concentratedand the residue purified by normal phase flash chromatography (40 gcolumn, eluting with 5% EtOAc in hexanes) to provide 870 mg (70%) of thetitle compound as a colorless oil. ¹H NMR (400 MHz. CDCl₃) δ 4.42 (d,J=10.2 Hz, 1H), 4.20 (q, J=7.4 Hz, 2H), 2.83 (m, 1H), 2.31 (s, 3H), 1.24(t, J=7.4 Hz, 3H), 1.09 (d, J=6.7 Hz, 3H), 0.82 (d, J=6.6 Hz, 3H); MS:(ES) m/z calculated for C₁₂H₁₇ClF₃N₂O₂[M+H]⁺ 313.1, found 313.1. Theregiochemistry of the system was confirmed by nOe between the α-methineproton and pyrazole methyl protons (and those of the isopropyl group).

b) The ester obtained above (870 mg, 2.8 mmol) was slurried in 8.3 mLTHF and 2.8 mL of 2 M LiOH (aq) at 50° C. After 80 minutes, thevolatiles were evaporated and 1 mL of 6 M HCl (aq) was added. Theresulting precipitate was collected, washed with 3×H₂O, and dried toafford the title compound as a white solid (844 mg, 94%). MS: (ES) m/zcalculated for C₁₀H₁₃ClF₃N₂O₂ [M+H]⁺ 285.1, found 285.0.

c) The title compound was prepared from2-(4-chloro-5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)-3-methylbutanoicacid and1-(4-fluorophenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyridine usingGeneral Method B. The product mixture was washed with 2×1 M NaHSO₄. Thereaction slurry was purified by flash chromatography (SiO₂, 24 g column,eluting with 5-50% EtOAc in hexanes) to provide 89 mg (74%) of the titlecompound as a glassy white solid. ¹H NMR (400 MHz, CDCl₃) δ 8.44 (s,1H), 7.44 (m, 2H), 7.16 (m, 2H), 5.03 (d, J=10.9 Hz, 1H), 3.89 (ddd,J=2.7, 8.2, 10.9 Hz, 1H), 3.78 (ddd, J=2.7, 7.8, 10.9 Hz, 1H), 2.96 (m,1H), 2.79 (m, 2H), 2.37 (s, 3H), 1.99 (m, 1H), 1.82 (m, 1H), 1.12 (d,J=6.3 Hz, 3H), 0.79 (d, J=7.1 Hz, 3H), MS: (ES) m/z calculated forC₂₂H23ClF₄N₅O [M+H]⁺ 484.1, found 484.1.

Example 26: Synthesis of1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-[3-(trifluoromethyl)pyrazolo[3,4-b]pyridin-1-yl]ethanone

a) Ethyl bromoacetate (1.336 g, 8 mmol),3-iodo-1H-pyrazolo[3,4-b]pyridine (1.953 mg, 8 mmol), and K₂CO₃ (2.211g, 16 mmol) were combined in 32 mL of 2:1 THF:DMF at 60° C. After 4hours, the volatiles were removed, the residue diluted with 50 mL ofEtOAc, and the slurry washed with 3×brine (half-saturated). The organiclayer was concentrated and the residue purified by flash chromatography(SiO₂, 80 g column, eluting with 5-20% EtOAc in hexanes) to provide2.172 g (82%) of the product as granular white solid. MS: (ES) m/zcalculated for C₁₀H₁₁N₃O₂ [M+H]⁺ 332.0, found 332.0.

b) Methyl fluorosulfonyldifluoroacetate (MFSDA, 1.25 mL, 10.5 mmol) wasadded dropwise to a slurry of ethyl2-(3-iodo-1H-pyrazolo[3,4-b]pyridin-1-yl)acetate (994 mg, 3 mmol), CuI(571 mg, 3 mmol), and DMF (30 mL). The mixture was heated to 100° C. for1 hour and diluted in 50 mL of EtOAc. The reaction mixture was filteredthrough a pad of Celite, washing with additional EtOAc. The filtrate wasconcentrated to half volume and an equal volume of brine was addedresulting in the formation of a precipitate. The solid was collected andwashed with H₂O and dried at 60° C. to provide the title compound as avoluminous crystalline solid (1.366 g, 87%).

c) The ester obtained above (546 mg, 2 mmol) was slurried in 6 mL THFand 1.5 mL of 2 M LiOH (aq) at 50° C. After completion, the volatileswere evaporated and 3 mL of 1 M NaHSO₄ (aq) was added. The resultingprecipitate was collected, washed with 3×H₂O, and dried to afford thetitle compound as a white powder (415 mg, 85%). MS: (ES) m/z calculatedfor C₉H₇F₃N₃O₂ [M+H]⁺ 246.0, found 246.0.

d) The title compound was prepared from2-(3-(trifluoromethyl)-1H-pyrazolo[3,4-b]pyridin-1-yl)acetic acid and1-(4-fluorophenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyridine usingGeneral Method B. The reaction slurry was purified by flashchromatography (SiO₂, 24 g column, eluting with 5-70% EtOAc in hexanes)to provide 58 mg of the title compound as a white solid. ¹H NMR (400MHz, CDCl₃, mixture of rotamers) δ 8.65 (dd, J=2.6, 4.7 Hz, 1H), 8.32(s, 0.8H), 8.24 (d, J=8.2 Hz, 1H), 7.72 (s, 0.2H), 7.44-7.52 (m, 2H),7.33 (dd, J=4.3, 8.2 Hz, 1H), 7.13-7.23 (m, 2H), 5.76 (s, 0.4H), 5.65(s, 1.6H), 3.93 (m, 2H), 2.89 (t, J=6.3 Hz, 2H), 2.17 (m, 1.6H), 2.06(m, 0.4H); MS: (ES) m/z calculated for C₂₁H₁₇F₄N₆O [M+H]⁺445.1, found445.1.

Example 27: Synthesis of1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-[3-(trifluoromethyl)-4,5,6,7-tetrahydropyrazolo[3,4-b]pyridin-1-yl]ethanone

1-[1-(4-Fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-[3-(trifluoromethyl)pyrazolo[3,4-b]pyridin-1-yl]ethanone(22.2 mg, 0.05 mmol) prepared above was dissolved in 1 mL of methanoland treated with ˜20 mg of PtO₂ in a septum-capped vial. The vial waspurged and filled with hydrogen gas twice and then placed under ahydrogen atmosphere (balloon) and stirred overnight. The slurry wasfiltered and the filtrate purified by reverse phase HPLC (C18 column,acetonitrile-H₂O with 0.1% TFA as eluent) to provide the title compound(solution was neutralized and extracted to provide the free base) as awhite solid (13 mg, 60%0). ¹H NMR (400 MHz, CDCl₃, mixture of rotamers)δ 8.36 (s, 0.85H), 7.71 (s, 0.15H), 7.43-7.49 (m, 2H), 7.14-7.19 (m,2H), 5.15 (s, 0.3H), 5.00 (s, 1.7H), 4.29 (br s, 1H), 3.88-3.93 (m, 2H),3.29 (m, 2H), 2.85 (t, 0.1=6.3 Hz, 2H), 2.63 (t, J=6.2 Hz, 2H), 2.09 (m,2H), 1.85 (m, 2H); MS: (ES) m/z calculated for C₂₁H₂₁F₄N₆O [M+H]⁺ 449.1,found 449.1.

Example 28: Synthesis of1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-[3-(trifluoromethyl)-5,6-dihydro-4H-pyrano[2,3-c]pyrazol-1-yl]ethanone

a) 3-(Trifluoromethyl)-1,4,5,6-tetrahydropyrano[2,3-c]pyrazole (96 mg,0.5 mmol), K₂CO₃ (885 mg, 6.4 mmol), and 8 mL of 3:1 THF:DMF werecombined and treated with 71 μL (0.64 mmol) of ethyl bromoacetate. Theslurry was stirred at 50° C. overnight and the volatiles evaporated. Theresidue was diluted in EtOAc and H₂O and the layers separated. Theorganic layer was washed with brine and concentrated. The residue waspurified by flash chromatography (SiO₂, 40 g column, eluting with 5-20%EtOAc in hexanes) to provide 70 mg (39%) of the compound as a clear oil.¹H NMR (400 MHz, CDCl₃) δ 4.73 (s, 2H), 4.31 (dd, J=5.1, 6.1 Hz, 2H),4.23 (q, J=7.0 Hz, 2H), 2.65 (t, J=6.7 Hz, 2H), 2.00 (m, 2H), 1.28 (t,J=7.0 Hz, 3H); MS: (ES) m/z calculated for C₁₁H₁₄F₃N₂O₃[M+H]⁺ 279.1,found 279.1.

b) The ester obtained above (70 mg, 0.25 mmol) was slurried in 1 mL THFand 0.5 mL of 2 M LiOH (aq) at 50° C. After 90 minutes, the volatileswere evaporated and 2 mL of 1 M NaHSO₄ (aq) was added. The mixture wasextracted with 2×EtOAc. The organic layer was dried on Na₂SO₄, filtered,and concentrated to a foamy white solid (45 mg, 71%). MS: (ES) m/zcalculated for C₉H₁₀F₃N₂O₃ [M+H]⁺ 251.1, found 251.1.

c) The title compound was prepared from2-(3-(trifluoromethyl)-5,6-dihydropyrano[2,3-c]pyrazol-1(4H)-yl)aceticacid and 1-(4-fluorophenyl)-4,5,6,7-tetrahydro-H-pyrazolo[4,3-b]pyridineusing General Method B. The reaction slurry was purified by reversephase HPLC (C18 column, acetonitrile-H₂O with 0.1% TFA as eluent) toprovide 32 mg (70%) of the title compound as a white foam. ¹H NMR (400MHz, CDCl₃, mixture of rotamers) δ 8.40 (s, 0.8H), 7.56 (s, 0.2H),7.43-7.48 (m, 2H), 7.13-7.21 (m, 2H), 5.12 (s, 0.4H), 5.00 (s, 1.6H),4.34 (dd, J=5.1, 5.1 Hz, 2H), 3.91 (m, 0.4H), 3.80 (m, 1.6H), 2.86 (t,J=6.2 Hz, 1.6H), 2.83 (t, J=6.2 Hz, 0.4H), 2.67 (t, J=6.2 Hz, 2H), 2.09(m, 1.6H), 2.00-2.05 (m, 2.4H); MS: (ES) m/z calculated for C₂₁H₂₀F₄N₅O₂[M+H]⁺ 450.1, found 450.1.

Example 29: Synthesis of2-[4-chloro-5-methyl-3-(trifluoromethyl)pyrazol-1-yl]-1-[1-(2,4-difluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]ethanone

a) To a 100-mL round-bottomed flask was added2,4-difluorophenylhydrazine hydrochloride (1.48 g, 8.2 mmol),3-fluoro-2-formylpyridine (1.00 g, 8 mmol), K₂CO₃ (3.32 g, 24 mmol), andDMF (10 mL). The slurry was heated to 120° C. in an oil bath for 16hours at which point the reaction mixture was cooled and poured into 25mL of H₂O. The brown precipitate was collected via filtration and washedwith 3×H₂O. The solid was dissolved in CH₂Cl₂, washed with brine, driedon Na₂SO₄, filtered, and concentrated to a brown solid. This solid wasdissolved in EtOAc, eluted through a silica plug, and concentrated to abrown solid (1.58 g, 85%). MS: (ES) m/z calculated for C₁₂H₅F₂N₃ [M+H]⁺232.1, found 232.1.

b) The heterocycle obtained above (1.58 g, 6.8 mmol) was dissolved in 34mL of methanol and treated with 20% Pd(OH)₂/C (1.00 g) in a 500-mL Parrbottle. The system was purged and filled with hydrogen (3×) and thenshaken under 55 psi hydrogen until the starting material was consumed.The bottle was flushed with nitrogen and the reaction mixture filteredthrough a pad of Celite, washing with several portions of methanol. Thefiltrate was concentrated to provide the product (1.10 g) as a lightorange-brown oil. MS: (ES) m/z calculated for C₁₂H₁₂F₂N₃ [M+H]⁺ 236.1,found 236.1.

c) The compound was prepared2-(4-chloro-5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)acetic acid and1-(2,4-difluorophenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyridineusing General Method B. The product mixture was washed with 2×1 MNaHSO₄. The organic layer was concentrated and the residue was purifiedby reverse phase HPLC (C18 column, acetonitrile-H₂O with 0.1% TFA aseluent) to provide the title compound as a white foamy solid (47 mg,70%). ¹H NMR (400 MHz, CDCl₃, mixture of rotamers) δ 8.39 (s, 0.8H),7.64 (s, 0.2H), 7.48 (m, 2H), 7.00 (m, 2H), 5.25 (s, 0.4H), 5.14 (s,1.6H), 3.92 (m, 0.4H), 3.85 (m, 1.6H), 2.70 (m, 2H), 2.33 (s, 2.4H),2.32 (s, 0.6H), 2.11 (m, 1.6H), 2.03 (m, 0.4H); MS: (ES) m/z calculatedfor C₁₉H₁₆ClF₅N₅O [M+H]⁺ 460.1, found 460.1.

Examples 30 and 31: Synthesis of (2R)- and(2S)-2-[4-chloro-5-methyl-3-(trifluoromethyl)pyrazol-1-yl]-1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]butan-1-one

a) 2-(4-Chloro-5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)butanoicacid (677 mg, 2.5 mmol) was dissolved in 10 mL of CHCl₃ and 3 drops ofDMF. Oxalyl chloride (262 μL, 3 mmol) was added dropwise, causing gasevolution. After 30 minutes, (R)-phenylglycinol (377 mg, 2.75 mmol) wasadded followed by 8 mL of saturated NaHCO₃. After 45 minutes, thereaction mixture was washed with brine and purified by flashchromatography (SiO₂, 40 g column, eluting with 10-70%/o EtOAc inhexanes). First eluting diastereomer: ¹H NMR (400 MHz, CDCl₃) δ 7.62 (d,J=7.8 Hz, 1H), 7.23-7.39 (m, 3H), 7.08-7.11 (m, 2H), 5.00-5.04 (m, 1H),4.66 (t, J=7.4 Hz, 1H), 3.84 (m, 2H), 2.31 (t, J=7.4 Hz, 3H), 2.26 (s,3H), 0.91 (t, J=7.0 Hz, 3H); MS: (ES) m/z calculated for C₁₇H₂₀ClF₃N₃O₂[M+H]⁺ 390.1, found 390.1. Second eluting diasteromer: ¹H NMR (400 MHz,CDCl₃) δ 7.58 (d, J=7.0 Hz, 1H), 7.38 (m, 2H), 7.32 (m, 1H), 7.23-7.25(m, 2H), 5.00 (dt, J=5.1, 7.0 Hz, 1H), 4.69 (t, J=7.8 Hz, 1H), 3.75 (t,J=5.5 Hz, 2H), 2.32 (s, 3H), 2.10-2.27 (m, 3H), 0.84 (t, J=7.4 Hz, 3H);MS: (ES) m/z calculated for C₁₇H₂₀ClF₃N₃O₂ [M+H]⁺ 390.1, found 390.1.

b) The diastereomers obtained above were independently heated to 80° C.in 0.25 M dioxane with 15 equiv of 6 M H₂SO₄. The reaction slurries wereconcentrated to approx half volume and extracted with 3×EtOAc. Theorganic layer was dried on Na₂SO₄, filtered, and concentrated to providethe desired acid products in 82-89% yield.

c)(2R)-2-[4-Chloro-5-methyl-3-(trifluoromethyl)pyrazol-1-yl]-1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]butan-1-oneand(2S)-2-[4-Chloro-5-methyl-3-(trifluoromethyl)pyrazol-1-yl]-1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]butan-1-one:The acid obtained from the first eluting diastereomer in step a wascoupled with1-(4-fluorophenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyridine usingGeneral Method A. The reaction slurry was purified by reverse phase HPLC(C18 column, acetonitrile-H₂O with 0.1% TFA as eluent) to provide 73 mg(62%) of the title compound as a white solid. ¹H NMR (400 MHz, CDCl₃) δ8.43 (s, 1H), 7.44 (m, 2H), 7.16 (m, 2H), 5.29 (dd, J=6.7, 8.6 Hz, 1H),3.81 (ddd, J=2.7, 7.4, 10.6 Hz, 1H), 3.50 (ddd, J=2.8, 8.6, 11.8 Hz,1H), 2.79 (dt, J=2.7, 6.3 Hz, 2H), 2.33 (m, 1H), 2.30 (s, 3H), 2.22 (m,1H), 1.96 (m, 1H), 1.86 (m, 1H), 0.98 (t, I=7.0 Hz, 3H); MS: (ES) m/zcalculated for C₂₁H₂₁ClF₄N₅O [M+H]⁺ 470.1, found 470.1. The acidobtained from the second eluting diastereomer in step a was coupled with1-(4-fluorophenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyridine usingGeneral Method A. The reaction slurry was purified by reverse phase HPLC(C18 column, acetonitrile-H₂O with 0.1% TFA as eluent) to provide 71 mg(60%) of the title compound as a white solid. ¹H NMR (400 MHz, CDCl₃) δ8.43 (s, 1H), 7.44 (m, 2H), 7.16 (m, 2H), 5.29 (dd, J=6.7, 8.6 Hz, 1H),3.81 (ddd, J=2.7, 7.4, 10.6 Hz, 1H), 3.50 (ddd, J=2.8, 8.6, 11.8 Hz,1H), 2.79 (dt, J=2.7, 6.3 Hz, 2H), 2.33 (m, 1H), 2.30 (s, 3H), 2.22 (m,1H), 1.96 (m, 1H), 1.86 (m, 1H), 0.98 (t, J=7.0 Hz, 3H); MS: (ES) m/zcalculated for C₂₁H₂₁ClF₄N₅O [M+H]⁺ 470.1, found 470.1.

Example 32: Synthesis of1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-[1-(trifluoromethyl)imidazo[1,5-α]pyridin-3-yl]ethanone

a) To ethyl 2-(imidazo[1,5-α]pyridin-3-yl)acetate (131 mg, 0.64 mmol) inCH₃CN was added 3,3-dimethyl-1-(trifluoromethyl)-1,2-benziodoxole (212mg, 0.64 mmol). The slurry was heated to 80° C. for 95 minutes thencooled to room temperature. The reaction mixture was purified by flashchromatography (SiO₂, 40 g column, eluting with 15-75% EtOAc in hexanes)to provide the product (46 mg, 26%) as an orange oil that solidifies onstanding. MS: (ES) m/z calculated for C₁₂H₁₂F₃N₂O₂[M+H]⁺ 273.1, found273.0.

b) The ester obtained above (36 mg, 0.13 mmol) was slurried in 1 mL THFand 0.125 mL of 2 M LiOH (aq) at 50° C. Upon completion, the volatileswere evaporated and 500 μL of 1 M NaHSO₄ (aq) and 4 mL of EtOAc wereadded. The layers were separated and the organic phase washed withbrine. The organic layer was dried on Na₂SO₄, filtered, and concentratedto a tan solid (29 mg, 90%). MS: (ES) m/z calculated for C₁₀H₈F₃N₂O₂[M+H]⁺ 245.1, found 245.0.

c) The title compound was prepared from2-(1-(trifluoromethyl)imidazo[1,5-α]pyridin-3-yl)acetic acid and1-(4-fluorophenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyridine usingGeneral Method A. The reaction slurry was purified by reverse phase HPLC(C18 column, acetonitrile-H₂O with 0.1%0/TFA as eluent) to provide 32 mg(72%) of the title compound as a white solid. ¹H NMR (400 MHz, CDCl₃,mixture of rotamers) δ 8.36 (d, J=6.6 Hz, 1H), 8.35 (s, 0.9H), 8.06 (s,0.1H), 7.63 (d, J=9.0 Hz, 1H), 7.50 (m, 0.3H), 7.42 (m, 1.7H), 7.12-7.22(m, 2H), 7.01 (m, 1H), 6.79 (t, J=6.6 Hz, 1H), 4.51 (s, 0.2H), 4.38 (s,1.8H), 4.08 (m, 1.7H), 3.90 (m, 0.3H), 2.86 (t, J=6.6 Hz, 0.3H), 2.80(t, J=6.6 Hz, 1.7H), 2.02 (m, 2H); MS: (ES) m/z calculated forC₂₂H₁₈F₄N₅O [M+H]⁺ 444.1, found 444.1.

Example 33: Synthesis of1-[1-(4-fluorophenyl)-5-methyl-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-[5-methyl-3-(trifluoromethyl)pyrazol-1-yl]butan-1-one

The title compound was prepared from2-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)butanoic acid and1-(4-fluorophenyl)-5-methyl-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyridineusing General Method A. The reaction slurry was purified by reversephase HPLC (C18 column, acetonitrile-H₂O with 0.1% TFA as eluent) toprovide 13 mg (46%) of the title compound as a white solid. ¹H NMR (400MHz, CDCl₃) 8.40 (s, 1H), 7.47 (m, 2H), 7.16 (m, 2H), 6.35 (s, 1H), 5.37(dd, J=6.7, 8.6 Hz, 1H), 4.63 (m, 1H), 2.86 (m, 1H), 2.73 (ddd, J=1.9,5.1, 11.3 Hz, 1H), 2.37 (s, 3H), 2.27 (m, 2H), 1.92-1.97 (m, 2H), 0.93(t, J=6.5 Hz, 3H), 0.57 (t, J=6.7 Hz, 3H); MS: (ES) m/z calculated forC₂₂H₂₄F₄N₅O [M+H]⁺ 450.2, found 450.2.

Example 34: Synthesis of1-[I-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-[3-(1-hydroxy-1-methyl-ethyl)-5-methyl-pyrazol-1-yl]butan-1-one

a) To a THF-rinsed 250-mL round-bottomed flask was added 80 mL of THFand 6.17 g (40 mmol) of ethyl 5-methyl-1H-pyrazole-3-carboxylate. Theflask was fitted with an addition funnel and cooled in an ice bath underN₂. Methyl magnesium bromide (3 M in Et₂O, 41 mL, 124 mmol) was addeddropwise. After 70 minutes, an additional 4 mL of Grignard reagent wasadded. At 2 hours, the reaction was quenched by the slow addition of 2mL of 1 M NaHSO₄.

The reaction slurry was acidified by the slow addition of 6 M HCl (aq).The volatiles were evaporated and the mixture extracted with 3×EtOAc.The organic layer was dried on MgSO₄, filtered, and concentrated toprovide a viscous yellow-orange oil (6.0 g, quant) that solidified onstanding. MS: (ES) m/z calculated for C₇H13N₂O [M+H]⁺ 141.1, found141.1.

b) Methyl bromobutyrate (1.068 g, 5.9 mmol),2-(5-methyl-1H-pyrazol-3-yl)propan-2-ol (752 mg, 5.36 mmol), andK₂CO₃(1.520 g, 11 mmol) were combined in 16 mL of 2:1 THF:DMF at 70° C.overnight. The reaction mixture was cooled and the solids were filteredoff washing with EtOAc. The filtrate was washed with 2×H₂O and brine.The organic layer was concentrated and the residue purified by flashchromatography (SiO₂, 80 g column, eluting with 10-65% EtOAc in hexanes)to provide 114 mg (9/o) of the title compound as a colorless oil. ¹H NMR(400 MHz, CDCl₃) 5.93 (s, 1H), 4.62 (dd, J=5.8, 9.8 Hz, 1H), 3.72 (s,3H), 2.95 (br s, 1H), 2.29 (m, 2H), 2.24 (s, 3H), 1.52 (s, 6H), 0.85 (t,J=7.4, 3H); MS: (ES) m/z calculated for C₁₂H₂₁N₂O₃ [M+H]⁺ 241.2, found241.2.

c) The ester obtained above (113 mg, 0.47 mmol) was slurried in 1 mL THFand 310 μL of 2 M LiOH at 50° C. After 30 minutes, the volatiles wereevaporated and 1 mL of 1 M NaHSO₄ was added. The slurry was extractedwith 3×EtOAc and the organic layer dried on Na₂SO₄, filtered, andconcentrated to give a clear oil (95 mg, 89%).

d) The title compound was prepared from2-(3-(2-hydroxypropan-2-yl)-5-methyl-1H-pyrazol-1-yl)butanoic acid and1-(4-fluorophenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyridine usingGeneral Method A. The reaction slurry was purified by reverse phase HPLC(C18 column, acetonitrile-H₂O with 0.1% TFA as eluent) to provide 3.7 mgof the title compound as an off-white solid. ¹H NMR (400 MHz, CDCl₃) δ8.46 (s, 1H), 7.44 (m, 2H), 7.15 (t, J=8.6 Hz, 2H), 5.92 (s, 1H), 5.17(dd, J=7.4, 7.4 Hz, 1H), 3.73 (m, 1H), 3.48 (m, 1H), 2.75 (m, 2H), 2.33(m, 1H), 2.23 (s, 3H), 2.17 (s, 1H), 2.14 (m, 1H), 1.87 (m, 1H), 1.73(m, 1H), 1.52 (s, 6H), 0.96 (t, J=7.4 Hz, 3H); MS: (ES) m/z calculatedfor C₂₃H₂₉FN₅O₂ [M+H]⁺ 426.2, found 426.2.

Example 35: Synthesis of1-[l-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-(3-isopropenyl-5-methyl-pyrazol-1-yl)butan-1-one

A portion of1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-[3-(1-hydroxy-1-methyl-ethyl)-5-methyl-pyrazol-1-yl]butan-1-onewas treated with 6 M HCl until complete. The reaction slurry waspurified by reverse phase HPLC (C18 column, acetonitrile-H₂O with 0.1%TFA as eluent) to provide 12 mg of the title compound as an off-whitesolid. ¹H NMR (400 MHz, CDCl₃) 8.47 (s, 1H), 7.44 (m, 2H), 7.14 (t,J=8.6 Hz, 2H), 6.14 (s, 1H), 5.42 (br s, 1H), 5.22 (dd, J=7.1, 8.2 Hz,1H), 5.02 (br s, 1H), 3.80 (ddd, J=3.1, 7.8, 10.1 Hz, 1H), 3.47 (ddd,J=2.8, 9.0, 12.1 Hz, 1H), 2.74 (m, 2H), 2.33 (m, 1H), 2.23 (s, 3H), 2.15(m, 1H), 2.10 (s, 3H) 1.87 (m, 1H), 1.78 (m, 1H), 0.98 (t, J=7.4 Hz,3H); MS: (ES) m/z calculated for C₂₃H₂₇FN₅O [M+H]⁺ 408.2, found 408.2.

Example 36: Synthesis of1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-[5-methyl-3-(trifluoromethyl)pyrazol-1-yl]propan-1-one

a) A mixture of potassium carbonate (922 mg, 6.67 mmol),2-methyl-5-(trifluoromethyl)-1H-pyrazole (500 mg, 3.33 mmol) and ethyl2-bromopropanoate (0.48 mL, 3.7 mmol) in dimethylformamide:tetrahydrofuran (3 mL: 6 mL) was heated at 60° C. for 5 h with stirring.After cooling to room temperature, most of tetrahydrofuran was removedby gently blowing nitrogen over the reaction mixture. Ethyl acetate andwater were added and the layers were separated. The aqueous layer wasextracted twice more with ethyl acetate. The combined organic layerswere dried (Na₂SO₄), filtered, and concentrated in vacuo. The cruderesidue was purified by flash chromatography (SiO₂, 10-20% ethyl acetatein hexanes) to afford the desired product (735 mg, 2.94 mmol, 88%).

b) Ethyl 2-[5-methyl-3-(trifluoromethyl)pyrazol-1-yl]propanoate (734 mg,2.94 mmol) was dissolved in tetrahydrofuran (4 mL) at room temperature.Aqueous lithium hydroxide solution (1.5 N, 2 mL, 3.0 mmol) was addedfollowed by methanol (˜2 mL) to make a uniform solution. After 2 h, mostof tetrahydrofuran and methanol was removed by gently blowing nitrogenover reaction mixture. Aqueous hydrochloric acid (5 N, 0.6 mL, 3.0 mmol)was added to the reaction mixture and the product was extracted withdichloromethane three times. The combined organic layers were dried overanhydrous sodium sulfate and concentrated under reduced pressure to givethe crude product (625 mg, 2.82 mmol, 96%) as a white solid which wasused without further purification.

c) Dimethylformamide (1 mL) was added to a mixture containing2-[5-methyl-3-(trifluoromethyl)pyrazol-1-yl]propanoic acid (49.6 mg,0.223 mmol) and HATU (89.1 mg, 0.234 mmol). Diisopropylethylamine (98μL, 0.56 mmol) was added at room temperature and the reaction mixturewas stirred for 1 minute.1-(4-Fluorophenyl)-4,5,6,7-tetrahydropyrazolo[4,3-b]pyridine (49.5 mg,0.228 mmol) was added all at once and the reaction mixture was stirredat room temperature overnight. Ethyl acetate and water were then addedand the layers were separated. The aqueous layer was extracted twicemore with ethyl acetate. The combined organic layers were dried(Na₂SO₄), filtered, and concentrated in vacuo. The crude residue waspurified by flash chromatography (SiO₂, 20-38% ethyl acetate in hexanes)to afford the desired product as a white solid (83.5 mg, 0.198 mmol,89%). ¹H NMR (400 MHz, CDCl₃) δ 8.45 (s, 1H), 7.44 (ddd, J=9.2, 5.2, 2.0Hz, 2H), 7.15 (dt, J=8.8, 2.0 Hz, 2H), 6.31 (s, 1H), 5.55 (q, J=7.2 Hz,1H), 3.73 (ddd, J=13, 7.6, 3.2 Hz, 1H), 3.37 (ddd, J=12, 8.8, 3.2 Hz,1H), 2.72-2.79 (m, 2H), 2.29 (s, 3H), 1.86-1.97 (m, 1H), 1.76-1.85 (m,1H), 1.80 (d, J=7.2 Hz, 3H); MS: (ES) m/z calculated for C₂₀H₁₉N₅OF₄[M+H]⁺ 422, found 422.

Example 37: Synthesis of1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-[5-methyl-3-(trifluoromethyl)pyrazol-1-yl]butan-1-one

a) A mixture of potassium carbonate (924 mg, 6.69 mmol),2-methyl-5-(trifluoromethyl)-1H-pyrazole (501 mg, 3.34 mmol) and methyl2-bromobutyrate (0.42 mL, 3.7 mmol) in dimethylformamide:tetrahydrofuran (3 mL: 6 mL) was heated at 60° C. for 5 h with stirring.After cooling to room temperature, most of tetrahydrofuran was removedby gently blowing nitrogen over the reaction mixture. Ethyl acetate andwater were added and the layers were separated. The aqueous layer wasextracted twice more with ethyl acetate. The combined organic layerswere dried (Na₂SO₄), filtered, and concentrated in vacuo. The cruderesidue was purified by flash chromatography (SiO₂, 12-17% ethyl acetatein hexanes) to afford the desired product (704 mg, 2.81 mmol, 84%).

b) Methyl 2-[5-methyl-3-(trifluoromethyl)pyrazol-1-yl]butyrate (703 mg,2.81 mmol) was dissolved in tetrahydrofuran (4 mL) at room temperature.Aqueous lithium hydroxide solution (1.5 N, 2 mL, 3.0 mmol) was addedfollowed by methanol (˜2 mL) to make a uniform solution. After 2 h, mostof tetrahydrofuran and methanol was removed by gently blowing nitrogenover reaction mixture. Aqueous hydrochloric acid (5 N, 0.6 mL, 3.0 mmol)was added to the reaction mixture and the mixture was extracted withdichloromethane three times. The combined organic layers were dried overanhydrous sodium sulfate and concentrated under reduced pressure whichyielded the crude product (661 mg, 2.80 mmol, 100%) as a white solidwhich was used without further purification.

c) Dimethylformamide (0.7 mL) was added to a mixture containing2-[5-methyl-3-(trifluoromethyl)pyrazol-1-yl]butyric acid (50.5 mg, 0.214mmol) and HATU (84.1 mg, 0.221 mmol). Diisopropylethylamine (92 μL, 0.53mmol) was added at room temperature and the reaction mixture was stirredfor 1 minute.1-(4-Fluorophenyl)-4,5,6,7-tetrahydropyrazolo[4,3-b]pyridine (44.1 mg,0.203 mmol) was then added all at once and the reaction mixture wasstirred at room temperature overnight. Ethyl acetate and water wereadded and the layers were separated. The aqueous layer was extractedtwice more with ethyl acetate. The combined organic layers were dried(Na₂SO₄), filtered, and concentrated in vacuo. The crude residue waspurified by flash chromatography (SiO₂, 17-33% ethyl acetate in hexanes)to afford the desired product as a white solid (80.9 mg, 0.186 mmol,92%). ¹H NMR (400 MHz, CDCl₃) δ 8.44 (s, 1H), 7.44 (ddd, J=8.8, 5.2, 2.0Hz, 2H), 7.15 (ddd, J=8.4, 8.4, 2.0 Hz, 2H), 6.30 (s, 1H), 5.32 (dd,J=8.4, 6.8 Hz, 1H), 3.78 (ddd, J=12, 7.2, 2.8 Hz, 1H), 3.47 (ddd, J=12,8.8, 3.2 Hz, 1H), 2.69-2.81 (m, 2H), 2.31 (s, 3H), 2.18-2.38 (m, 2H),1.88-1.97 (m, 1H), 1.74-1.83 (m, 1H), 0.98 (t, J=7.2 Hz, 3H); MS: (ES)min calculated for C₂₁H₂₁N₅OF₄ [M+H]⁺ 436, found 436.

Example 38: Synthesis of1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-[2-methyl-4-(trifluoromethyl)imidazol-1-yl]propan-1-one

a) A mixture of potassium carbonate (832 mg, 6.02 mmol),2-methyl-5-(trifluoromethyl)-1H-imidazole (450 mg, 3.00 mmol) and ethyl2-bromopropanoate (0.43 mL, 3.3 mmol) in dimethylformamide:tetrahydrofuran (2 mL: 4 mL) was heated at 50° C. for 7 h with stirringand then at room temperature overnight. Most of the tetrahydrofuran wasremoved by gently blowing nitrogen over the reaction mixture. Ethylacetate and water were then added and the layers were separated. Theaqueous layer was extracted twice more with ethyl acetate. The combinedorganic layers were dried (Na₂SO₄), filtered, and concentrated in vacuo.The crude residue was purified by flash chromatography (SiO₂, 17-50%ethyl acetate in hexanes) to afford the desired product (697 mg, 2.79mmol, 93%).

b) Ethyl 2-[2-methyl-4-(trifluoromethyl)imidazol-1-yl]propanoate (697mg, 2.79 mmol) was dissolved in tetrahydrofuran (5 mL) at roomtemperature. Aqueous lithium hydroxide solution (1.5 N, 2.5 mL, 3.8mmol) was added followed by methanol (˜2 mL) to make a uniform solution.After 2 h, most of tetrahydrofuran and methanol were removed by gentlyblowing nitrogen over reaction mixture. Aqueous hydrochloric acid (5 N,0.75 mL, 3.8 mmol) was added to the reaction mixture and the product wasextracted with dichloromethane three times. The combined organic layerswere dried over anhydrous sodium sulfate and concentrated under reducedpressure which yielded the crude product (562 mg, 2.52 mmol, 91%) aswhite foam which was used without further purification.

c) 2-[2-Methyl-4-(trifluoromethyl)imidazol-1-yl]propanoic acid (151 mg,0.679 mmol) was dissolved in dichloromethane (2 mL) at room temperature.Oxalyl chloride (120 μL, 1.38 mmol) and a catalytic amount ofdimethylformamide (1 μL) was added and the reaction was stirred at roomtemperature for 2 h. Solvent was removed under reduced pressure and theacid chloride was dried under vacuum. The crude acid chloride wasdissolved in dichloromethane (2.1 mL). To a portion of this solution(0.7 mL, 0.22 mmol) was added1-(4-fluorophenyl)-4,5,6,7-tetrahydropyrazolo[4,3-b]pyridine (41.5 mg,0.191 mmol) and pyridine (55 μL, 0.68 mmol) at room temperature and thereaction mixture was stirred for 2 h. Water was added and the layerswere separated. The aqueous layer was extracted twice more withdichloromethane.

The combined organic layers were dried (Na₂SO₄), filtered, andconcentrated in vacuo. The crude residue was purified by flashchromatography (SiO₂, 40-100% ethyl acetate in hexanes with 0.1% aqueousammonium hydroxide) to afford the desired product as a white solid (70.1mg, 0.166 mmol, 87%). H NMR (400 MHz, CDCl₃) δ 8.44 (s, 1H), 7.45 (ddd,J=9.2, 4.8, 2.0 Hz, 2H), 7.17 (dt, J=9.2, 2.0 Hz, 2H), 5.21 (q, J=6.8Hz, 1H), 3.75 (ddd, J=12, 7.2, 3.2 Hz, 1H), 3.51 (ddd, J=12, 8.0, 3.2Hz, 1H), 2.84 (t, J=6.6 Hz, 2H), 2.47 (s, 3H), 1.96-2.09 (m, 2H), 1.72(d, J=6.4 Hz, 3H); MS: (ES) m/z calculated for C₂₀H₁₉N₅OF₄ [M+H]⁺ 422,found 422.

Example 39: Synthesis of1-[l-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-[2-methyl-4-(trifluoromethyl)imidazol-1-yl]butan-1-one

a) A mixture of potassium carbonate (834 mg, 6.04 mmol),2-methyl-5-(trifluoromethyl)-1H-imidazole (449 mg, 2.99 mmol) and methyl2-bromobutyrate (0.38 mL, 3.3 mmol) in dimethylformamide:tetrahydrofuran (2 mL: 4 mL) was heated at 50° C. for 7 h with stirringand then at room temperature overnight. Most of the tetrahydrofuran wasthen removed by gently blowing nitrogen over the reaction mixture. Ethylacetate and water were added and the layers were separated. The aqueouslayer was extracted twice more with ethyl acetate. The combined organiclayers were dried (Na₂SO₄), filtered, and concentrated in vacuo. Thecrude residue was purified by flash chromatography (SiO₂, 20-50° % ethylacetate in hexanes) to afford the desired product (641 mg, 2.56 mmol,86%).

b) Methyl 2-[2-methyl-4-(trifluoromethyl)imidazol-1-yl]butyrate (641 mg,2.56 mmol) was dissolved in tetrahydrofuran (5 mL) at room temperature.Aqueous lithium hydroxide solution (1.5 N, 2.2 mL, 3.3 mmol) was addedfollowed by methanol (˜2 mL) to make a uniform solution. After 2 h, mostof tetrahydrofuran and methanol was removed by gently blowing nitrogenover the reaction mixture. Aqueous hydrochloric acid (5 N, 0.66 mL, 3.3mmol) was added to the reaction mixture and the product was extractedwith dichloromethane three times. The combined organic layers were driedover anhydrous sodium sulfate and concentrated under reduced pressurewhich yielded the crude product (497 mg, 2.11 mmol, 82%) as a white foamwhich was used without further purification.

c) 2-[2-Methyl-4-(trifluoromethyl)imidazol-1-yl]butyric acid (149 mg,0.632 mmol) was dissolved in dichloromethane (2 mL) at room temperature.Oxalyl chloride (110 μL, 1.26 mmol) and a catalytic amount ofdimethylformamide (1 μL) was added and the reaction was stirred at roomtemperature for 2 h. Solvent was removed under reduced pressure and theacid chloride was dried under vacuum. The crude acid chloride wasdissolved in acetonitrile (1.5 mL). To a portion of this solution (0.5mL, 0.21 mmol) was added1-(4-fluorophenyl)-4,5,6,7-tetrahydropyrazolo[4,3-b]pyridine (38.7 mg,0.178 mmol) and pyridine (55 μL, 0.68 mmol) at room temperature and thereaction mixture was stirred for 2 h. Water and ethyl acetate were thenadded and the layers were separated. The aqueous layer was extractedtwice more with ethyl acetate. The combined organic layers were dried(Na₂SO₄), filtered, and concentrated in vacuo. The crude residue waspurified by flash chromatography (SiO₂, 50-80% ethyl acetate in hexaneswith 0.1% aqueous ammonium hydroxide) to afford the desired productcontaminated with some2-[2-methyl-4-(trifluoromethyl)imidazol-1-yl]butyric acid. The productwas dissolved in ethyl acetate and washed once with 1N aqueous sodiumhydroxide solution. Drying over anhydrous sodium sulfate followed byfiltration and removal of solvent under reduced pressure and dryingunder vacuum afforded the desired product as a white solid (60.4 mg,0.139 mmol, 78%). ¹H NMR (400 MHz, CDCl₃) δ 8.20 (s, 1H), 7.84 (d, J=1.2Hz, 1H), 7.58 (ddd, J=9.2, 5.2, 2.0 Hz, 2H), 7.35 (ddd, J=9.2, 8.4, 0.8Hz, 2H), 5.47 (dd, J=9.2, 5.6 Hz, 1H), 3.81-3.98 (m, 2H), 2.84 (t, J=6.2Hz, 2H), 2.34 (s, 3H), 2.03-2.11 (m, 2H), 1.96-2.01 (m, 2H), 0.85 (t,J=7.2 Hz, 3H); MS: (ES) m/z calculated for C₂₁H₂₁N₅OF₄ [M+H]⁺ 436, found436.

Example 40: Synthesis of1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-[4-(trifluoromethoxy)phenyl]ethanone

To a mixture of1-(4-fluorophenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyridine (0.023g, 0.10 mmol), 4-(trifluoromethoxy)phenylacetic acid (0.025 g, 0.11mmol) and Et₃N (0.060 mL, 0.43 mmol) in DMF (0.6 mL) was added HATU(0.060 g, 0.15 mmol). The mixture was stirred at rt for 30 min. It wasthen quenched with water. The mixture was partitioned between EtOAc (50mL) and sat. NaHCO₃ (30 mL). The organic layer was separated, dried overNa₂SO₄, concentrated in vacuo, and purified by reverse phase HPLC toafford the desired product (0.040 g, 95%). ¹H NMR (400 MHz, CDCl₃) δ8.51 (s, 1H), 7.43 (m, 2H), 7.30 (m, 2H), 7.18 (m, 4H), 3.91 (s, 2H),3.76 (m, 2H), 2.79 (t, J=6.4 Hz, 2H), 2.00 (m, 2H); MS: (ES) m/zcalculated for C₂₁H₁₇F₄N₃O₂ [M+H]⁺ 420.1, found 420.1.

Example 41: Synthesis of2-[4-chloro-3-(trifluoromethoxy)phenyl]-1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]ethanone

To a mixture of1-(4-fluorophenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyridine (0.023g, 0.10 mmol), 4-chloro-3-(trifluoromethoxy)phenylacetic acid (0.025 g,0.10 mmol) and Et₃N (0.060 mL, 0.43 mmol) in DMF (0.6 mL) was added HATU(0.060 g, 0.15 mmol). The mixture was stirred at rt for 30 min. It wasthen quenched with water and partitioned between EtOAc (50 mL) and sat.NaHCO₃ (30 mL). The organic layer was separated, dried over Na₂SO₄,concentrated in vacuo and purified by reverse phase HPLC to afford thedesired product (0.039 g, 85%). ¹H NMR (400 MHz, CDCl₃) δ 8.47 (s, 1H),7.44 (m, 3H), 7.12-7.22 (m, 4H), 3.90 (s, 2H), 3.76 (m, 2H), 2.80 (t,J=6.2 Hz, 2H), 2.02 (m, 2H); MS: (ES) m/z calculated for C₂₁H₁₆ClF₄N₃O₂[M+H]⁺ 454.1, found 454.1.

Example 42: Synthesis of2-[4-chloro-3-(trifluoromethyl)phenyl]-1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-methyl-propan-1-one

To a mixture of1-(4-fluorophenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyridine (0.030g, 0.11 mmol), 2-[4-chloro-3-(trifluoromethyl)phenyl]-2-methyl-propanoicacid (0.030 g, 0.11 mmol) and NEt₃ (0.070 mL, 0.50 mmol) in DMF (0.6 mL)was added HATU (0.060 g, 0.15 mmol). The mixture was stirred at rt for30 min. It was then quenched with water, and partitioned between EtOAc(50 mL) and sat. NaHCO₃ (30 mL). The organic layer was separated, driedover Na₂SO₄, concentrated in vacuo and purified by reverse phase HPLC toafford the desired product (0.005 g, 10%). ¹H NMR (400 MHz, CDCl₃) 8.50(s, 1H), 7.60 (d, J=2.0 Hz, 1H), 7.30-7.51 (m, 4H), 7.14 (dd, J=8.8, 8.4Hz, 2H), 3.22 (m, 2H), 2.69 (t, J=6.4 Hz, 2H), 1.64 (s, 6H); 1.62 (m,2H); MS: (ES) m/z calculated for C₂₃H₂₀ClF₄N₃O [M+H]⁺ 466.1, found466.1.

Example 43: Synthesis of2-(4-chloropyrazol-1-yl)-1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]ethanone

A mixture of 2-[(4-chloro-pyrazol-1-yl)]acetic acid (0.050 g, 0.31mmol), (COCl)₂ (0.060 mL, 0.70 mmol) and DMF (1 drop) in CH₂Cl₂ (1 mL)was stirred for 30 min at rt. It was then evaporated to dryness on ahigh vacuum pump. The obtained oil was added to a mixture containing1-(4-fluorophenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyridine (0.020g, 0.092 mmol) and NEt₃ (0.070 mL, 0.50 mmol) in CH₂Cl₂ (1 mL). Theresulting mixture was then stirred at rt for 30 min. It was thenquenched with water and partitioned between EtOAc (50 mL) and sat.NaHCO₃ (30 mL). The organic layer was separated, dried over Na₂SO₄,concentrated in vacuo, and purified by reverse phase HPLC to afford thedesired product (0.012 g, 36%). ¹H NMR (400 MHz, CDCl₃) δ 8.41 (s, 1H),7.60 (s, 1H), 7.51 (s, 1H), 7.44 (dd, J=8.8, 4.6 Hz, 2 H), 7.17 (dd,J=9.2, 8.8 Hz, 2H), 5.16 (s, 2H), 3.82 (m, 2H), 2.85 (t, 0.1=6.2 Hz,2H), 2.09 (m, 2H); MS: (ES) m/z calculated for C₁₇H₁₅ClFN₅O [M+H]⁺360.1, found 360.1.

Example 44: Synthesis of2-[4-chloro-3-(difluoromethyl)-5-methyl-pyrazol-1-yl]-1-[I-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]ethanone

To a mixture of1-(4-fluorophenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyridine (0.023g, 0.10 mmol), 2-(4-chloro-3-difluoromethyl-5-methyl-pyrazol-1-yl)aceticacid (0.023 g, 0.10 mmol) and NEt₃ (0.065 mL, 0.46 mmol) in DMF (0.6 mL)was added HATU (0.060 g, 0.15 mmol). The mixture was stirred at rt for30 min. It was then quenched with water and partitioned between EtOAc(50 mL) and sat. NaHCO₃ (30 mL). The organic layer was separated, driedover Na₂SO₄, concentrated in vacuo and purified by reverse phase HPLC toafford the desired product (0.032 g, 75%). ¹H NMR (400 MHz, CDCl₃) δ8.36 (s, 1H), 7.44 (m, 2H), 7.16 (dd, J=8.8, 8.4 Hz, 2H), 6.67 (d, J=54Hz, 1H), 5.11 (s, 2H), 3.83 (m, 2H), 2.85 (t, J=6.2 Hz, 2H), 2.30 (s,3H), 2.10 (m, 2H); MS: (ES) m/z calculated for C₁₉H₁₇ClF₃N₅O [M+H]⁺424.1, found 424.1.

Example 45: Synthesis of2-[4-chloro-3-(trifluoromethyl)pyrazol-1-yl]-1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]ethanone

a) A mixture of 3-(trifluoromethyl)pyrazole (1.58 g, 11.6 mmol) and NCS(1.55 g, 11.6 mmol) in CH₃CN (20 mL) was heated at 80° C. for 3 hrs. Itwas then cooled to rt, evaporated in vacuo, and purified by flashchromatography (SiO₂) with a gradient elution of 0 to 15% EtOAc/CH₂Cl₂to afford 4-chloro-3-(trifluoromethyl)pyrazole (1.52 g, 77%).

b) A mixture of 4-chloro-3-(trifluoromethyl)pyrazole (0.026 g, 0.15mmol),2-chloro-1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridine-4-yl]ethanone(0.045 g, 0.15 mmol) and K₂CO₃(0.043 g, 0.31 mmol) in THF (0.6 mL) andDMF (0.3 mL) was heated at 60° C. for 1 hr. It was then cooled to rt.The mixture was partitioned between EtOAc (50 mL) and sat. NaHCO₃ (30mL). The organic layer was separated, dried over Na₂SO₄, concentrated invacuo and purified by flash chromatography on SiO₂ with a gradientelution of 0-60% EtOAc/hexanes to afford the desired product (0.056 g,87%). ¹H NMR (400 MHz, CDCl₃) δ 8.37 (s, 1H), 7.70 (s, 1H), 7.45 (m,2H), 7.16 (dd, J=8.4, 8.4 Hz, 2H), 5.17 (s, 2H), 3.81 (m, 2H), 2.87 (t,J=6.2 Hz, 2H), 2.12 (m, 2H); MS: (ES) m/z calculated for C₁₈H₁₄ClF₄N₅O[M+H]⁺ 428.1, found 428.1.

Example 46: Synthesis of1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-[3-(trifluoromethyl)pyrazol-1-yl]ethanone

To a mixture of1-(4-fluorophenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyridine (0.026g, 0.12 mmol), 2-(3-(trifluoromethyl)-pyrazol-1-yl)acetic acid (0.024 g,0.12 mmol) and NEt₃ (0.060 mL, 0.43 mmol) in DMF (0.5 mL) was added HATU(0.060 g, 0.15 mmol). The mixture was stirred at rt for 30 min. It wasthen quenched with water. The mixture was partitioned between EtOAc (50mL) and sat. NaHCO₃ (30 mL). The organic layer was separated, dried overNa₂SO₄, concentrated in vacuo and purified by reverse phase HPLC toafford the desired product (0.035 g, 74%). ¹H NMR (400 MHz, CDCl₃) δ8.38 (s, 1H), 7.64 (m, 1H), 7.43 (m, 2H), 7.15 (dd, J=8.8, 8.4 Hz, 2H),6.62 (d, J=2.4 Hz, 1H), 5.23 (s, 2H), 3.82 (m, 2H), 2.84 (t, J=6.4 Hz,2H), 2.08 (m, 2H); MS: (ES) m/z calculated for C₁₈H₁₅F₄N₅O [M+H]⁺394.1,found 394.1.

Example 47: Synthesis of2-(4-chloro-5-methyl-pyrazol-1-yl)-1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]ethanone

a) A mixture of 2-(5-methylpyrazol-1-yl)acetic acid (0.700 g, 5 mmol)and NCS (0.668 g, 5 mmol) in CH₃CN (10 mL) was heated at 80° C. for 1.5hrs. It was then cooled to rt, evaporated in vacuo, and purified byflash chromatography (SiO₂, with a gradient elution of 0-20%MeOH/CH₂Cl₂) to afford 2-(4-chloro-5-methylpyrazol-1-yl)acetic acid(0.870 g, 100%).

b) To a mixture of1-(4-fluorophenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyridine (0.026g, 0.12 mmol), 3-(trifluoromethyl)-pyrazol-1-yl)acetic acid (0.024 g,0.12 mmol) and NEt₃ (0.060 mL, 0.43 mmol) in DMF (0.5 mL) was added HATU(0.060 g, 0.15 mmol). The mixture was stirred at rt for 30 min. It wasthen quenched with water. The mixture was partitioned between EtOAc (50mL) and sat. NaHCO₃ (30 mL). The organic layer was separated, dried overNa₂SO₄, concentrated in vacuo and purified by reverse phase HPLC toafford the desired product (0.025 g, 55%). ¹H NMR (400 MHz, CDCl₃) δ8.40 (s, 1H), 7.48 (s, 1H), 7.44 (dd, J=8.8, 4.6 Hz, 2H), 7.16 (dd,J=8.4, 8.4 Hz, 2H), 5.16 (s, 2H), 3.84 (m, 2H), 2.84 (t, J=6.4 Hz, 2H),2.29 (s, 3H), 2.10 (m, 2H); MS: (ES) m/z calculated for C₁₅H₁₇ClFN₅O[M+H]⁺ 374.1, found 374.1.

Example 48: Synthesis of1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-[4-(trifluoromethyl)pyrazol-1-yl]ethanone

A mixture of 4-(trifluoromethyl)-1H-pyrazole (0.030 g, 0.22 mmol),2-chloro-1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridine-4-yl]ethanone(0.025 g, 0.085 mmol) and K₂CO₃ (0.060 g, 0.43 mmol) in THF (0.8 mL) andDMF (0.4 mL) was heated at 65° C. for 1 hr. It was then cooled to rt.The mixture was partitioned between EtOAc (50 mL) and sat. NaHCO₃ (30mL). The organic layer was separated, dried over Na₂SO₄, concentrated invacuo and purified by reverse phase HPLC to afford the desired product(0.038 g, 100%). ¹H NMR (400 MHz, CDCl₃) δ 8.40 (s, 1H), 7.89 (s, 1H),7.77 (s, 1H), 7.44 (dd, J=8.8, 4.6 Hz, 2H), 7.16 (dd, J=8.8, 8.4 Hz,2H), 5.20 (s, 2H), 3.83 (m, 2H), 2.85 (t, J=6.2 Hz, 2H), 2.10 (m, 2H),MS: (ES) m/z calculated for C₁₈H₁₅F₄N₅O [M+H]⁺ 394.1, found 394.1.

Example 49: Synthesis of1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-[3-(trifluoromethyl)-6,7-dihydro-4H-pyrano[4,3-c]pyrazol-1-yl]ethanone

A mixture of 3-(trifluoromethyl)-1,4,6,7-tetrahydropyrano[4,3-c]pyrazole(0.050 g, 0.26 mmol),2-chloro-1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridine-4-yl]ethanone(0.050 g, 0.17 mmol) and K₂CO₃ (0.130 g, 0.94 mmol) in THF (0.8 mL) andDMF (0.4 mL) was heated at 55° C. for 1 hr. It was then cooled to rt.The mixture was partitioned between EtOAc (50 mL) and sat. NaHCO₃ (30mL). The organic layer was separated, dried over Na₂SO₄, concentrated invacuo, and purified by reverse phase HPLC to afford the desired product(0.035 g, 55%). ¹H NMR (400 MHz, CDCl₃) δ 8.38 (s, 1H), 7.44 (dd, J=8.8,4.6 Hz, 2H), 7.17 (dd, J=8.8, 8.0 Hz, 2H), 5.13 (s, 2H), 4.75 (s, 2H),3.98 (t, J=5.4 Hz, 2H), 3.87 (m, 2H), 2.85 (t, J=6.2 Hz, 2H), 2.79 (t,J=5.6 Hz, 2H), 2.10 (m, 2H); MS: (ES) m/z calculated for C₂₁H₁₉F₄N₅O₂[M+H]⁺ 450.1, found 450.1.

Example 50: Synthesis ofi-[I-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-[4-(1,2,4-oxadiazol-3-yl)pyrazol-1-yl]ethanone

a) To a mixture of1-(4-fluorophenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyridine (0.400g, 1.83 mmol), 2-(4-iodo-pyrazol-1-yl)acetic acid (0.554 g, 2.2 mmol)and NEt₃ (0.642 mL, 4.59 mmol) in DMF (5 mL) was added HATU (0.836 g,2.2 mmol). The mixture was stirred at rt for 40 min. It was thenquenched with water. The mixture was partitioned between EtOAc (100 mL)and sat. NaHCO₃ (50 mL). The organic layer was separated, dried overNa₂SO₄, concentrated in vacuo, and purified by flash chromatography(SiO₂, with a gradient elution of 0˜30% EtOAc/CH₂Cl₂) to yield1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-(4-iodopyrazol-1-yl)ethanone(0.82 g, 100%).

b) A mixture of1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-(4-iodopyrazol-1-yl)ethanone(0.388 g, 0.86 mmol), Zn(CN)₂ (0.151 g, 1.29 mmol), Pd₂(dba)₃ (0.079 g,0.086 mmol) and dppf (0.072 g, 0.13 mmol) in DMF (10 mL) was heated at90° C. for 1 hr. It was then cooled to rt. The mixture was partitionedbetween EtOAc (10 mL) and sat. NaHCO₃ (50 mL). The organic layer wasseparated, dried over Na₂SO₄, concentrated in vacuo, and purified byflash chromatography with a gradient elution of 0-100% EtOAc/CH₂Cl₂ toafford2-(4-cyanopyrazol-1-yl)-1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]ethanone(0.205 g, 68%).

c) A mixture of2-(4-cyanopyrazol-1-yl)-1-[l-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]ethanone(0.175 g, 0.50 mmol), NH₂OH—HCl (0.500 g, 7.2 mmol) and NEt₃ (1.00 mL,7.1 mmol) in EtOH (3 mL) was heated at 90° C. for 5 hrs. It was thencooled to it. The mixture was partitioned between IPA/CHCl₃ (1:2, 50 mL)and sat. NaHCO₃ (30 mL). The organic layer was separated, dried overNa₂SO₄ and concentrated in vacuo to obtain1-[2-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-oxo-ethyl]-N′-hydroxy-pyrazole-4-carboxamidine(0.115 g, 60%).

d) A mixture of1-[2-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-oxo-ethyl]-N′-hydroxy-pyrazole-4-carboxamidine(0.115 g, 0.30 mmol) and p-TsOH.H₂O (0.030 g, 0.15 mmol) in HC(OMe)₃ (3mL) was heated at 100° C. for 1.5 hrs. It was then cooled to rt. Themixture was partitioned between IPA/CHCl₃ (1:2, 50 mL) and sat. NaHCO₃(30 mL). The organic layer was separated, dried over Na₂SO₄,concentrated in vacuo and purified by reverse phase HPLC to afford thedesired product (0.055 g, 21%). ¹H NMR (400 MHz, CDCl₃) δ 8.71 (s, 1H),7.65-8.50 (m, 3H), 7.44 (dd, J=8.4, 4.6 Hz, 2H), 7.17 (dd, J=9.2, 7.6Hz, 2H), 5.27 (m, 2H), 3.85 (m, 2H), 2.85 (m, 2H), 2.14 (m, 2H); MS:(ES) m/z calculated for C₁₉H₁₆FN₇O₂ [M+H]⁺ 394.1, found 394.1.

Example 51: Synthesis of1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-[3-(1H-imidazol-2-yl)pyrazolo[3,4-b]pyridin-1-yl]ethanone

To a mixture of1-(4-fluorophenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyridine (0.050g, 0.23 mmol),2-[3-(1H-imidazol-2-yl)pyrazolo[3,4-b]pyridine-1-yl]acetic acid (0.050g, 0.21 mmol) and Et₃N (0.15 mL, 1.07 mmol) in DMF (0.7 mL) was addedHATU (0.130 g, 0.33 mmol). The mixture was stirred at rt for 30 min. Itwas then quenched with water. The mixture was partitioned betweenIPA/CHCl₃ (1:2, 100 mL) and sat. NaHCO₃ (40 ml). The organic layer wasseparated, dried over Na₂SO₄, concentrated in vacuo, and purified byreverse phase HPLC to afford the desired product (0.090 g, 96%). ¹H NMR(TFA salt) (400 MHz, CD₃OD) δ 8.73 (dd, J=4.8, 1.6 Hz, 1H), 8.69 (dd,J=8.0, 1.6 Hz, 1H), 8.14 (s, 1H), 7.72 (s, 2H), 7.55 (m, 3H), 7.28 (dd,J=9.2, 8.4 Hz, 2H), 5.89 (s, 2H), 4.06 (m, 2H), 2.92 (t, J=6.2 Hz, 2H),2.20 (m, 2H); MS: (ES) m/z calculated for C₂₃H₁₉F₈O [M+H]⁺ 443.1, found443.1.

Example 52: Synthesis of1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-[3-(trifluoromethyl)-4,5,6,7-tetrahydroindazol-1-yl]ethanone

To a mixture of1-(4-fluorophenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyridine (0.023g, 0.11 mmol),2-[3-(trifluoromethyl)-4,5,6,7-tetrahydroindazol-1-yl]acetic acid (0.027g, 0.11 mmol) and NEt₃ (0.050 mL, 0.36 mmol) in DMF (0.6 mL) was addedHATU (0.050 g, 0.13 mmol). The mixture was stirred at rt for 30 min. Itwas then quenched with water. The mixture was partitioned betweenIPA/CHCl₃ (1:2, 50 mL) and sat. NaHCO₃ (30 mL). The organic layer wasseparated, dried over Na₂SO₄, concentrated in vacuo, and purified byreverse phase HPLC to afford the desired product (0.027 g, 55%). ¹H NMR(400 MHz, CDCl₃) δ 8.43 (s, 1H), 7.42 (dd, J=8.8, 4.4 Hz, 2H), 7.17 (dd,J=9.2, 8.0 Hz, 2H), 5.15 (s, 2H), 3.87 (m, 2H), 2.82 (t, J=6.2 Hz, 2H),2.61 (m, 4H), 2.11 (m, 2H), 1.87 (m, 2H), 1.79 (m, 2H); MS: (ES) m/zcalculated for C₂₂H₂₁F₄N₅O [M+H]⁺ 448.1, found 448.1.

Example 53: Synthesis of2-(1,3-benzoxazol-2-yl)-1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]ethanone

To a mixture of1-(4-fluorophenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyridine (0.023g, 0.11 mmol), benzooxazol-2-yl-acetic acid (0.023 g, 0.13 mmol) andNEt₃ (0.060 mL, 0.43 mmol) in DMF (0.6 mL) was added HATU (0.050 g, 0.13mmol). The mixture was stirred at rt for 30 min. It was then quenchedwith water. The mixture was partitioned between IPA/CHCl₃ (1:2, 50 mL)and sat. NaHCO₃ (30 mL). The organic layer was separated, dried overNa₂SO₄, concentrated in vacuo, and purified by reverse phase HPLC toafford the desired product (0.022 g, 53%). ¹H NMR (400 MHz, CDCl₃) δ8.46 (s, 1H), 7.71 (m, 1H), 7.53 (m, 1H), 7.45 (dd, J=8.8, 4.8 Hz, 2H),7.34 (m, 2H), 7.15 (dd, J=8.8, 8.4 Hz, 2H), 4.25 (s, 2H), 3.88 (m, 2H),2.83 (t, J=6.4 Hz, 2H), 2.08 (m, 2H); MS: (ES) m/z calculated forC₂₁H₁₇FN₄O₂ [M+H]⁺ 377.1, found 377.1.

Example 54: Synthesis of2-[4-chloro-5-methyl-3-(trifluoromethyl)pyrazol-1-yl]-1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-3-methoxy-propan-1-one

To a mixture of2-[4-chloro-5-methyl-3-(trifluoromethyl)pyrazol-1-yl]-1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]ethanone(0.045 g, 0.10 mmol) and NaH (0.030 g, 0.75 mmol, 60% in mineral oil) inDMF (0.8 mL) was added dry paraformaldehyde (0.015 g, 0.50 mmol). Themixture was stirred at rt for 30 min. It was then quenched with water.The mixture was partitioned between EtOAc (50 mL) and brine (30 mL). Theorganic layer was separated, dried over Na₂SO₄, concentrated in vacuo,and purified by reverse phase HPLC to afford the desired product (0.006g, 12%). ¹H NMR (400 MHz, CDCl₃) δ 8.44 (s, 1H), 7.44 (dd, J=8.8, 4.6Hz, 2H), 7.16 (dd, J=8.4, 8.4 Hz, 2H), 5.51 (dd, J=8.0, 7.2 Hz, 1H),4.18 (m, 1H), 4.08 (m, 1H), 3.75 (m, 1H), 3.37 (s, 3H), 3.35 (m, 1H),2.78 (m, 2H), 2.33 (s, 3H), 2.09 (m, 2H); MS: (ES) m/z calculated forC₂₁H₂₀ClF₄N₅O₂ [M+H]⁺ 486.1, found 486.1.

Example 55: Synthesis of2-[4-chloro-5-methyl-3-(trifluoromethyl)pyrazol-1-yl]-1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-3-hydroxy-propan-1-one

To a mixture of2-[4-chloro-5-methyl-3-(trifluoromethyl)pyrazol-1-yl]-1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]ethanone(0.100 g, 0.23 mmol) in THF (3 ml) at −78° C. was added LDA (0.15 mL,0.3 mmol, 2 M in THF). After stirred for 10 min at −78° C., dryparaformaldehyde (0.020 g, 0.66 mmol, suspended in THF) was added. Themixture was then warmed to rt for 8 min, quenched with sat. NH₄Cl. Themixture was partitioned between EtOAc (50 mL) and brine (30 mL). Theorganic layer was separated, dried over Na₂SO₄, concentrated in vacuo,and purified by reverse phase HPLC to yield the desired product (0.060g, 56%). ¹H NMR (400 MHz, CDCl₃) δ 8.45 (s, 1H), 7.43 (dd, J=8.4, 4.6Hz, 2H), 7.17 (dd, J=8.8, 8.4 Hz, 2H), 5.40 (t, J=5.4 Hz, 1H), 4.90 (s,1H), 4.30 (m, 1H), 4.20 (m, 1H), 3.71 (m, 1H), 3.29 (m, 1H), 2.79 (m,2H), 2.28 (s, 3H), 1.95 (m, 2H); MS: (ES) m/z calculated forC₂₀H₁₅ClF₄N₅O₂ [M+H]⁺ 472.1, found 472.1.

Example 56: Synthesis of1-[i-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-3-hydroxy-2-[5-methyl-3-(trifluoromethyl)pyrazol-1-yl]propan-1-one

To a mixture of1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-[5-methyl-3-(trifluoromethyl)pyrazol-1-yl]ethanone(0.100 g, 0.25 mmol) in THF (3 mL) at −78° C. was added LDA (0.15 mL,0.3 mmol, 2 M in THF). After stirring for 10 min at −78° C., dryparaformaldehyde (0.025 g, 0.82 mmol, suspended in THF) was added. Themixture was then warmed to rt for 8 min, followed by the addition ofsat. NH₄Cl. The mixture was partitioned between EtOAc (50 mL) and brine(30 mL). The organic layer was separated, dried over Na₂SO₄,concentrated in vacuo, and purified by reverse phase HPLC to yield thedesired product (0.056 g, 55%). ¹H NMR (400 MHz, CDCl₃) δ 8.50 (s, 1H),7.42 (m, 2H), 7.18 (dd, J=8.4, 8.4 Hz, 2H), 6.36 (s, 1H), 5.41 (t, J=5.6Hz, 1H), 4.36 (m, 1H), 4.20 (m, 1H), 3.69 (m, 1H), 3.26 (m, 1H), 2.76(m, 2H), 2.31 (s, 3H), 1.92 (m, 2H); MS: (ES) min calculated forC₂₀H₁₉F₄N₅O₂ [M+H]⁺ 438.1, found 438.1.

Example 57: Synthesis of1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-4-hydroxy-4-methyl-2-[5-methyl-3-(trifluoromethyl)pyrazol-1-yl]pentan-1-one

a) To a mixture of1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-[5-methyl-3-(trifluoromethyl)pyrazol-1-yl]ethanone(0.150 g 0.37 mmol) in THF (4 ml) at −78° C. was added LDA (0.247 mL,0.50 mmol, 2 M in THF). After stirring for 10 min at −78° C., ethylbromoacetate (0.061 mL, 0.55 mmol) was added. The mixture was thenwarmed to rt for 10 min and quenched with sat. NH₄Cl. The mixture waspartitioned between EtOAc (50 mL) and brine (30 mL). The organic layerwas separated, dried over Na₂SO₄, concentrated in vacuo and purified byflash chromatography (SiO₂ with a gradient elution of 0-60%EtOAc/hexanes) to afford ethyl4-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-3-[5-methyl-3-(trifluoromethyl)pyrazol-1-yl]-4-oxo-butanoate(0.120 g, 83%).

b) A mixture of ethyl4-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-3-[5-methyl-3-(trifluoromethyl)pyrazol-1-yl]-4-oxo-butanoate(0.028 g, 0.10 mmol) and MeMgCl (0.100 mL, 0.30 mmol, 3 M in THF) in THF(1 mL) was stirred at rt for 10 min. It was then quenched with sat.NH₄Cl and partitioned between EtOAc (50 mL) and brine (30 mL). Theorganic layer was separated, dried over Na₂SO₄, concentrated in vacuoand purified by reverse phase HPLC to yield the desired product (0.005g, 17%). ¹H NMR (400 MHz, CDCl₃) δ 8.45 (s, 1H), 7.43 (m, 2H), 7.16 (dd,J=8.8, 8.4 Hz, 2H), 6.31 (s, 1H), 5.72 (t, J=6.4 Hz, 1H), 3.88 (m, 1H),3.52 (m, 1H), 2.75 (m, 3H), 2.32 (s, 3H), 2.25 (m, 2H), 1.96 (m, 1H),1.85 (m, 1H), 1.34 (s, 3H), 1.18 (s, 3H); MS: (ES) m/z calculated forC₂₃H₂₅F₄N₅O₂[M+H]⁺ 480.1, found 480.1.

Example 58: Synthesis of1-[i-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-[5-methyl-3-(trifluoromethyl)pyrazol-1-yl]-3-tetrahydropyran-4-yl-propan-1-one

A mixture of1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-[5-methyl-3-(trifluoromethyl)pyrazol-1-yl]ethanone(0.100 g, 0.25 mmol), 4-(iodomethyl)tetrahydro-2H-pyran (0.165 g, 0.75mmol) and NaH (0.030 g, 0.75 mmol, 60% in mineral oil) in DMF (0.8 mL)was stirred at rt for 1.5 hrs. It was then quenched with sat. NH₄Cl. Themixture was partitioned between EtOAc (50 mL) and brine (30 mL). Theorganic layer was separated, dried over Na₂SO₄, concentrated in vacuo,and purified by reverse phase HPLC to afford the desired product (0.007g, 6%). ¹H NMR (400 MHz, CDCl₃) δ 8.44 (s, 1H), 7.43 (dd, J=8.8, 4.8 Hz,2H), 7.16 (dd, J=8.8, 8.4 Hz, 2H), 6.31 (s, 1H), 5.54 (m, 1H), 3.95 (m,2H), 3.78 (m, 1H), 3.47 (m, 1H), 3.34 (m, 2H), 2.75 (m, 2H), 2.30 (s,3H), 2.20 (m, 3H), 1.88 (m, 3H), 1.40 (m, 3H); MS: (ES) m/z calculatedfor C₂₅H₂₇F₄N₅O₂ [M+H]⁺ 506.2, found 506.2.

Example 59: Synthesis of4-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-3-[5-methyl-3-(trifluoromethyl)pyrazol-1-yl]-4-oxo-butanamide

a) A mixture of ethyl4-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-3-[5-methyl-3-(trifluoromethyl)pyrazol-1-yl]-4-oxo-butanoate(0.030 g, 0.060 mmol), LiOH*H₂O (0.020 g, 0.47 mmol), THF (0.4 mL), MeOH(0.4 mL) and H₂O (0.2 mL) was stirred for 30 min at rt. It was thenacidified with 1 M aq. HCl (2 mL) and extracted with IPA/CHCl₃ (1:2, 50mL). The organic layer was separated, dried over Na₂SO₄, andconcentrated in vacuo to yield4-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-3-[5-methyl-3-(trifluoromethyl)pyrazol-1-yl]-4-oxo-butanoicacid (0.029 g, 1000).

b) To a mixture of4-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-3-[5-methyl-3-(trifluoromethyl)pyrazol-1-yl]-4-oxo-butanoicacid (0.029 g, 0.08 mmol) and NH₃ (0.3 mL, sat. in CH₂Cl₂) in DMF (0.6mL) was added HATU (0.040 g, 0.10 mmol). The mixture was stirred at rtfor 30 min. It was then quenched with water. The mixture was partitionedbetween IPA/CHCl₃ (1:2, 50 mL) and sat. NaHCO₃ (30 mL). The organiclayer was separated, dried over Na₂SO₄, concentrated in vacuo, andpurified by reverse phase HPLC to afford the desired product (0.014 g,37%). ¹H NMR (400 MHz, CDCl₃) δ 8.44 (s, 1H), 7.42 (dd, J=9.2, 4.6 Hz,2H), 7.16 (dd, J=8.4, 8.0 Hz, 2H), 6.53 (s, 1H), 6.34 (s, 1H), 6.32 (s,1H), 5.92 (m, 1H), 3.80 (m, 1H), 3.41 (m, 1H), 3.33 (m, 1H), 3.00 (m,1H), 2.76 (m, 2H), 2.37 (s, 3H), 1.98 (m, 1H), 1.88 (m, 1H); MS: (ES)m/z calculated for C₂₁H₂₀F₄N₆O₂ [M+H]⁺ 465.1, found 465.1.

Example 60: Synthesis of methyl4-[2-[4-chloro-5-methyl-3-(trifluoromethyl)pyrazol-1-yl]acetyl]-1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazol[4,3-b]pyridine-5-carboxylate

a) To a cooled solution of 1-(4-fluorophenyl)pyrazolo[4,3-b]pyridine (1g, 4.67 mmol) in CH₂Cl₂ (50 mL) under nitrogen atmosphere was addedmCPBA (75%, 1.2 g, 5.14 mmol) in one portion. The resulting solution wasallowed to warm to room temperature and stirred for 12 h. The reactionmixture was then diluted with CH₂Cl₂ (100 mL) and washed with saturatedNaHCO₃ solution (100 mL). The aqueous layer was extracted with CH₂Cl₂(50 mL). The combined organic layers were dried (MgSO₄), filtered andconcentrated in vacuo. The crude residue was purified by flashchromatography (SiO₂, 20% MeOH in CH₂Cl₂) to afford the desired product(1.1 g, 4.7 mmol, quantitative yield).

b) To a solution of1-(4-fluorophenyl)-4-oxido-pyrazolo[4,3-b]pyridin-4-ium (916 mg, 4 mmol)in CH₃CN (10 mL) was added TMSCN (800 μL, 6 mmol) and Et₃N (556 μL, 4mmol) and stirred at 80° C. overnight. The reaction mixture was thencooled to room temperature and concentrated in vacuo. The crude residuewas purified by flash chromatography (SiO₂, 80%/EtOAc in hexanes) togive the desired product (857 mg, 3.6 mmol, 90%).

c) Water (10 mL) was added to1-(4-fluorophenyl)pyrazolo[4,3-b]pyridine-5-carbonitrile (600 mg, 2.52mmol) and the mixture was cooled to 0° C. followed by the addition ofconcentrated H₂SO₄ (10 mL) slowly drop wise. The obtained yellowishclear solution was then stirred at 110° C. for 12 h. The reactionmixture was cooled to 0° C. followed by the slow addition of 10 N NaOHdrop wise with stirring until reached pH 5-6. The resulting white solidwas filtered and washed with water (30 mL) and heptanes (50 mL), and wasthen dried under high vacuum to obtain the desired crude product (700mg) which was used in the next step without further purification.

d) To 1-(4-fluorophenyl)pyrazolo[4,3-b]pyridine-5-carboxylic acid (650mg, 2.5 mmol) was added CH₂Cl₂ (40 mL) and MeOH (5 mL) followed byTMSCHN₂ (2 M in Et₂O, 10 mL, excess) drop wise. The resulting yellowsuspension was stirred at room temperature for an hour. The reactionmixture was then diluted with CH₂Cl₂ (40 mL), washed with saturatedNaHCO₃ solution (50 mL), dried (MgSO₄) and concentrated in vacuo toobtain the desired crude product (750 mg) which was used directly in thenext step without further purification.

e) MeOH (6 mL) and 12 N HCl (1 mL) were added to methyl1-(4-fluorophenyl)pyrazolo[4,3-b]pyridine-5-carboxylate (100 mg, 0.37mmol) followed by 20% Pd(OH)₂ on carbon (100 mg, excess) and theresulting mixture was stirred under H₂ gas (55 psi) on a Parr shaker atroom temperature for 36 h. The reaction mixture was then filteredthrough a small pad of celite, washed with MeOH (20 mL) and concentratedin vacuo. The obtained residue was dissolved in EtOAc (50 mL) and washedwith saturated NaHCO₃ solution (20 mL), dried (MgSO₄) and concentratedin vacuo to obtain the desired crude product (30 mg) which was used assuch in next step without further purification.

f) To a solution of2-[4-chloro-5-methyl-3-(trifluoromethyl)pyrazol-1-yl]acetic acid (20 mg,0.073 mmol) in DMF (2 mL) was added Et₃N (50 μL, 0.219 mmol), HATU (55mg, 0.145 mmol) followed by methyl1-(4-fluorophenyl)-4,5,6,7-tetrahydropyrazolo[4,3-b]pyridine-5-carboxylate(20 mg, 0.073 mmol) at room temperature. The resulting solution was thenstirred at 80° C. for 5 h. The reaction mixture was then cooled to roomtemperature, H₂O (10 mL) was added and the mixture was extracted withEtOAc (2×15 mL). The combined EtOAc layers were dried (MgSO₄), filtered,concentrated in vacuo and the resulting crude product was purified byreverse phase HPLC (C18 column, acetonitrile-H₂O with 0.1% TFA aseluent) to give the desired product as white powder (0.004 g, 0.008mmol, 7%). ¹H NMR (400 MHz, CD₃OD) δ 8.28 (s, 1H), 7.52 (2H, dd, 1=5, 9Hz), 7.26 (2H, t, 8.6, J=17.2 Hz), 5.3 (2H, dd, J=17.2, 69.2 Hz), 3.83(s, 3H), 2.80-2.82 (m, 2H), 2.29 (s, 3H), 2.15-2.17 (m, 2H); MS: (ES)m/z calculated for C₂₁H₁₈ClF₄N₅O₃ [M+H]⁺ 500.1, found 500.1.

Example 61: Synthesis of1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazol[4,3-b]pyridin-4-yl]-2-[3-(trifluoromethyl)-5,6-dihydro-4H-cyclopenta[c]pyrazol-1-yl]ethanone

To a solution2-[3-(trifluoromethyl)-5,6-dihydro-4H-cyclopenta[c]pyrazol-1-yl]aceticacid (83 mg, 0.35 mmol) in DMF (2 mL) was added Et₃N (75 μL, 0.525mmol), HATU (160 mg, 0.42 mmol), followed by1-(4-fluorophenyl)-4,5,6,7-tetrahydropyrazolo[4,3-b]pyridine (77 mg,0.35 mmol) at room temperature. The resulting solution was then stirredat 50° C. for 30 min.

The reaction mixture was then cooled to room temperature, H₂O (10 mL)was added and the mixture was extracted with EtOAc (2:20 mL). Thecombined EtOAc layers were dried (MgSO₄), filtered, and concentrated invacuo. Purification by reverse phase HPLC (C18 column, acetonitrile-H₂Owith 0.1% TFA as eluent) to give the desired product as a white powder(0.011 g, 0.025 mmol, 7%). ¹H NMR (400 MHz, DMSO-d₆) δ 8.15 (s, 1H), 7.6(m, 2H), 7.35 (t, 2H, J=8.6, 17.6 Hz), 5.38 (s, 2H), 3.77 (m, 2H), 2.85(t, 2H, J=6.3, 12.5 Hz), 2.66 (m, 4H), 2.53 (m, 2H), 1.97 (m, 2H); MS:(ES) m/z calculated for C₂₁H₁₉F₄N₅O [M+H]⁺ 434.2, found 434.1.

Example 62: Synthesis of2-[4-chloro-3-(trifluoromethyl)phenyl]-1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]propan-1-one

a) To a solution of 4-(bromomethyl)-1-chloro-2-(trifluoromethyl)benzene(3.94 g, 14.4 mmol) in DMSO (29 mL) was added NaCN (1.06 g. 21.6 mmol).The mixture was heated at 50° C. for 1 h with stirring. After cooling toroom temperature, the reaction mixture was poured into a 100 mL beakercontaining ice water (25 mL). The aqueous layer was extracted withdichloromethane (4×10 mL), and the combined organic layers were washedwith brine and dried (Na₂SO₄), filtered, and concentrated in vacuo togive the desired product (2.5 g, 79%).

b) To a mixture of 2-[4-chloro-3-(trifluoromethyl)phenyl]acetonitrile(2.0 g, 9.1 mmol) and methyl iodide (1.29 g. 9.1 mmol) in toluene (20mL) at 80° C. was slowly added NaNH₂ (428 mg, 11 mmol). The mixture wasstirred at 80° C. for 1 hour. After cooling to room temperature, water(10 mL) was added. The mixture was extracted with ethyl acetate (2×15mL). The combined organic layers were washed with brine and dried(Na₂SO₄), filtered, and concentrated in vacuo. The crude residue waspurified by flash chromatography (SiO₂, 0-15% ethyl acetate in hexanes)to afford the desired product (500 mg, 24%).

c) To a solution of 2-[4-chloro-3-(trifluoromethyl)phenyl]propanenitrile(450 mg, 1.93 mmol) in dioxane (6 mL) was added sulfuric acid (60%, 6mL). The mixture was heated at 110° C. for 14 hours. After cooling toroom temperature, the reaction mixture was partitioned between water anddichloromethane. The aqueous layer was extracted with dichloromethane(2×10 mL), and the combined organic layers were washed with brine anddried (Na₂SO₄), filtered, and concentrated in vacuo to give the desiredproduct (350 mg, 72%).

d) To a mixture of 2-[4-chloro-3-(trifluoromethyl)phenyl]propanoic acid(47 mg, 0.184 mmol) and1-(4-fluorophenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyridine (40 mg,0.184 mmol) in DMF (1 mL) was added Hunig's base (59 mg, 0.46 mmol) and2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uraniumhexafluorophosphate methanaminium (HATU) (77 mg, 0.2 mmol). The mixturewas stirred at room temperature for 1 hour, and then was partitionedbetween water (4 mL) and ethyl acetate (6 mL). The organic layer waswashed with brine and dried (Na₂SO₄), filtered, concentrated in vacuo,and purified by reverse phase HPLC (C18 column, acetonitrile-H₂O with0.1% TFA as eluent) to give the desired product as a white solid (24 mg,29%0). ¹H NMR (400 MHz, CDCl₃) δ 8.51 (s, 1H), 7.62 (d, J=2.0 Hz, 1H),7.48-7.42 (m, 4H), 7.16-7.11 (m, 2H), 4.11 (m, 1H), 3.76 (m, 1H), 3.52(m, 1H), 2.77 (m, 2H), 1.96 (m, 1H), 1.80 (m, 1H), 1.55 (d, J=7.2 Hz,3H); MS: (ES) m/z calculated for C₂₂H₁₈ClF₄N₃ O [M+H]⁺ 452.1, found452.1.

Example 63: Synthesis of2-[2-chloro-4-(trifluoromethyl)phenyl]-1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]ethanone

To a mixture of 2-[2-chloro-4-(trifluoromethyl)phenyl]acetic acid (59mg, 0.23 mmol) and1-(4-fluorophenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyridine (50 mg,0.23 mmol) in DMF (1 mL) was added Hunig's base (59 mg, 0.46 mmol) and2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uraniumhexafluorophosphate methanaminium (HATU) (96 mg, 0.2 mmol). The mixturewas stirred at room temperature for 1 hour, and then was partitionedbetween water (4 mL) and ethyl acetate (6 mL). The organic layer waswashed with brine and dried (Na₂SO₄), filtered, concentrated in vacuo,and purified by reverse phase HPLC (C18 column, acetonitrile-H₂O with0.1% TFA as eluent) to give the desired product as a white solid (25 mg,25%). ¹H NMR (400 MHz, CDCl₃) δ 8.44 (s, 0.8H), 7.70 (s, 1H), 7.56-7.44(m, 4H), 7.38 (s, 0.2H), 7.18-7-14 (m, 2H), 4.14 (s, 2H), 3.82 (m, 2H),2.85 (m, 2H), 2.08 (m, 2H); MS: (ES) m/z calculated for C₂₁H₁₆ClF₄N₃ O[M+H]⁺ 438.1, found 438.1.

Example 64: Synthesis of2-[1H-benzimidazo-2-yl)-1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]ethanone

To a mixture of (1H-benzoimidazol-2yl)acetic acid (41 mg, 0.23 mmol) and1-(4-fluorophenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyridine (50 mg,0.23 mmol) in DMF (1 mL) was added Hunig's base (59 mg, 0.46 mmol) and2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uraniumhexafluorophosphate methanaminium (HATU, 96 mg, 0.2 mmol). The mixturewas stirred at room temperature for 1 hour and then was partitionedbetween water (4 mL) and ethyl acetate (6 mL). The organic layer waswashed with brine and dried (Na₂SO₄), filtered, concentrated in vacuo,and purified by reverse phase HPLC (C18 column, acetonitrile-H₂O with0.1% TFA as eluent) to give the desired product as a white solid (0.020g, 23%). ¹H NMR (400 MHz, CDCl₃) δ 8.38 (s, 0.8H), 8.00 (s, 1H), 7.85(m, 1H), 7.61 (s, 0.2H), 7.52-7.43 (m, 2H), 7.36-7-29 (m, 3H), 7.23-7.14(m, 2H), 5.18 (s, 2H), 3.84 (m, 2H), 2.88 (m, 2H), 2.13 (m, 2H); MS:(ES) m/z calculated for C₂₁H₁₆ClF₄N₃ O [M+H]⁺ 376.1, found 376.1.

Example 65: Synthesis of1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-pyrrolo[2,3-b]pyridin-1-yl-ethanone

a) A mixture of 1H-pyrrolo[2,3-b]pyridine (471.2 mg, 4.0 mmol), ethyl2-bromoacetate (530 μL, 4.8 mmol) and potassium carbonate (667.9 mg, 4.8mmol) in DMF (5 mL) was heated at 60° C. for 2 hours with stirring.After cooling to room temperature, the reaction mixture was diluted with20 ml of water, and extracted with EtOAc (2×30 mL). The combined organiclayer was then washed with water (2×20 mL), dried (Na₂SO₄), filtered,and concentrated in vacuo. The crude residue was purified by flashchromatography (SiO₂, 40% ethyl acetate in hexanes) to afford thedesired product (371.5 mg, 1.8 mmol, 45%).

b) A mixture of ethyl 2-pyrrolo[2,3-b]pyridin-1-ylacetate (371.5 mg, 1.8mmol), and lithium hydroxide (87.9 mg, 3.6 mmol) in a mixture of THF (4mL) and H₂O (1 mL) was stirred at room temperature overnight. Thereaction mixture was concentrated in vacuo, and acidified to pH 5 with 3N aqueous hydrochloric acid. The resulting solid was filtered, andwashed with H₂O (2×10 mL), and dried in vacuo to afford the desiredproduct (200.2 mg, 1.1 mmol, 62%).

c) A mixture of 2-pyrrolo[2,3-b]pyridin-1-ylacetic acid (51.2 mg, 0.24mmol), 1-(4-fluorophenyl)-4,5,6,7-tetrahydropyrazolo[4,3-b]pyridine(50.3 mg, 0.29 mmol), triethyl amine (200 μL, 1.4 mmol), and HATU (110.7mg, 0.29 mmol) in DMF (1 mL) was stirred at room temperature for 2hours. The reaction mixture was then diluted with 10 mL of aqueoussaturated sodium bicarbonate, and extracted with EtOAc (2×15 mL). Thecombined organic layers were then washed with water (2×20 mL), dried(Na₂SO₄), filtered, and concentrated in vacuo. The crude residue waspurified by flash chromatography (SiO₂, 70% ethyl acetate in hexanes) toafford the desired product (25.9 mg, 0.07 mmol, 28%). ¹H NMR (400 MHz,CDCl₃) δ 8.40 (s, 0.8H), 8.30 (dd, J=4.8, 1.6 Hz, 1H), 7.94 (dd, J=7.2,1.6 Hz, 1H), 7.81 (s, 0.2H), 7.45 (m, 2H), 7.36 (d, J=3.6 Hz, 0.8H),7.31 (d, J=3.6 Hz, 0.2H), 7.18 (m, 2H), 7.09 (dd, J=7.6, 4.8 Hz, 1H),6.58 (d, J=3.6 Hz, 1H), 5.44 (s, 0.2H), 5.32 (s, 0.8H), 3.91 (m, 2H),2.84 (m, 2H), 2.07 (m, 2H); MS: (ES) m/z calculated for C₂₁H₁₈FNO₅[M+H]⁺ 376.1, found 376.1.

Example 66: Synthesis of1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-imidazo[4,5-b]pyridin-3-yl-ethanone

a) A mixture of 3H-imidazo[4,5-b]pyridine (952.7 mg, 8.0 mmol), ethyl2-bromoacetate (1.0 mL, 9.6 mmol) and potassium carbonate (1.37 g, 9.6mmol) in DMF (10 mL) was heated at 60° C. for 2 hours with stirring.After cooling to room temperature, the reaction mixture was diluted with20 mL of water, and extracted with EtOAc (2×30 mL). The combined organiclayers were then washed with water (2×20 mL), dried (Na₂SO₄), filtered,and concentrated in vacuo. The crude residue was purified by flashchromatography (SiO₂, 100% ethyl acetate to 10% MeOH in EtOAc) to affordthe desired product (489.3 mg, 2.4 mmol, 30%).

b) A mixture of ethyl 2-imidazo[4,5-b]pyridin-3-ylacetate (400.3 mg, 1.9mmol), and lithium hydroxide (96.7 mg, 4.0 mmol) in a mixture of THF (4mL) and H₂O (1 ml) was stirred at room temperature overnight. Thereaction mixture was concentrated in vacuo, and acidified to pH 5 with 3N aqueous hydrochloric acid. The aqueous solution was then dried invacuo to afford the crude product, which was used directly in the nextstep.

c) A mixture of 2-imidazo[4,5-b]pyridin-3-ylacetic acid (251.7 mg,excess), 1-(4-fluorophenyl)-4,5,6,7-tetrahydropyrazolo[4,3-b]pyridine(42.7 mg, 0.23 mmol), triethyl amine (200 μL, 1.4 mmol), and HATU (87.9mg, 0.23 mmol) in DMF (1 mL) was stirred at room temperature for 2hours. The reaction mixture was diluted with 10 mL of aqueous saturatedsodium bicarbonate, and extracted with EtOAc (2×15 mL). The combinedorganic layers were then washed with water (2×20 mL), dried (Na₂SO₄),filtered, and concentrated in vacuo. The crude residue was purified byflash chromatography (SiO₂, 10% MeOH in EtOAc) to afford the desiredproduct (26.1 mg, 0.07 mmol, 30%). ¹H NMR (400 MHz, CDCl₃) δ 8.38 (d,J=7.2 Hz 1H), 8.50 (s, 0.8H), 8.25 (s, 0.2H), 8.12 (d, J=5.6 Hz, 1H),7.45 (m, 2H), 7.28 (m, 1H), 7.15 (m, 2H),), 5.46 (s, 0.2H), 5.33 (s,0.8H), 3.95 (m, 2H), 2.88 (m, 2H), 2.15 (m, 2H); MS: (ES) m/z calculatedfor C₂₀H₁₇FN₆O [M+H]⁺ 377.1, found 377.1.

Example 67: Synthesis of2-[4-chloro-5-methyl-3-(methylsulfonyl)pyrazol-1-yl]-1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]ethanone

a) To a mixture of1-(4-fluorophenyl)-4,5,6,7-tetrahydro-H-pyrazolo[4,3-b]pyridine (303 mg,1.40 mmol), 2-[4-chloro-5-methyl-3-(trifluoromethyl)pyrazol-1-yl]aceticacid (420 mg, 1.40 mmol), and i-Pr₂NEt (1 mL, 7.0 mmol) in DMF (5 mL)was added HATU (583 mg, 1.54 mmol). The resulting solution was thenstirred at room temperature overnight. The reaction mixture was thendiluted with EtOAc (50 mL) and washed with water (3×10 mL). The EtOAclayer was dried (Na₂SO₄), filtered, and concentrated in vacuo. The crudeproduct (480 mg, 70%) was used directly in the next step.

b) A mixture of2-(4-chloro-5-methyl-3-iodopyrazol-1-yl)-1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]ethanone(85 mg, 0.17 mmol), NaSO₂Me (52 mg, 0.51 mmol), CuI (98 mg, 0.51 mmol)in DMSO (3 mL) was heated at 110° C. overnight. After cooling to roomtemperature, the reaction mixture was diluted with EtOAc (50 mL) andwashed with water (3×10 mL). The EtOAc layer was dried (Na₂SO₄),filtered, concentrated in vacuo and purified by reverse phase HPLC (C18column, acetonitrile-H₂O with 0.1% TFA as eluent) to give the desiredproduct as white powder (42 mg, 55%). ¹H NMR (400 MHz, DMSO-d₆) 8.15 (s,1H), 7.60 (m, 2H), 7.38 (dd, J=8.9, 8.1 Hz, 2H), 5.58 (s, 2H), 3.80 (m,2H), 3.28 (s, 3H), 2.83 (t, J=5.7 Hz, 2H), 2.22 (s, 3H), 2.00 (m, 2H);MS: (ES) m/z calculated for C₁₉H₂₀ClFN₅O₃S [M+H]⁺ 452.1, found 452.0.

Example 68: Synthesis of2-(4-chloro-5-methyl-3-cyanopyrazol-1-yl)-1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]ethanone

A mixture of2-(4-chloro-5-methyl-3-iodopyrazol-1-yl)-1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]ethanone(430 mg, 0.86 mmol), Zn(CN)₂ (151 mg, 1.3 mmol), dppf (72 mg, 0.14mmol), and Pd₂(dba)₃ (79 mg, 0.09 mmol) in DMF (10 mL) and water (0.5mL) was heated at 90° C. overnight. After cooling to room temperature,the reaction mixture was diluted with EtOAc (30 mL) and washed withwater (3×5 mL). The EtOAc layer was dried (Na₂SO₄), filtered, andconcentrated in vacuo. The residue was triturated with 1:1 CH₂Cl₂/MeOH(3×5 mL) to give the desired product as white powder (250 mg, 72%). ¹HNMR (400 MHz, DMSO-d₆) δ 8.15 (s, 1H), 7.58 (m, 2H), 7.35 (dd, J=8.7,8.1 Hz, 2H), 5.58 (s, 2H), 3.80 (m, 2H), 2.84 (t, J=5.4 Hz, 2H), 2.25(s, 3H), 2.00 (m, 2H); MS: (ES) m/z calculated for C₁₉H₁₇ClF₄N₅O [M+H]⁺399.1, found 399.1.

Example 69: Synthesis of2-[4-chloro-3-(4,5-dihydro-1H-imidazol-2-yl)-5-methyl-pyrazol-1-yl]-1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]ethanonetrifluoroacetic acid salt

A mixture of2-(4-chloro-5-methyl-3-cyanopyrazol-1-yl)-1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]ethanone(150 mg, 0.38 mmol), ethylene diamine (2 mL) and acetic acid (0.3 mL) inethanol (5 mL) was heated at 100° C. for 2 hours. After cooling to roomtemperature, the reaction mixture was diluted with CH₂Cl₂ (50 mL) andwashed with water (3×10 mL). The organic layer was dried (Na₂SO₄),filtered, and concentrated in vacuo. Purification by reverse phase HPLC(C18 column, acetonitrile-H₂O with 0.1% TFA as eluent) gave2-[4-chloro-3-(4,5-dihydro-1H-imidazol-2-yl)-5-methyl-pyrazol-1-yl]-1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]ethanonetrifluoroacetic acid salt as white powder (84 mg, 51%). ¹H NMR (400 MHz,DMSO-d₆) δ 10.21 (s, 2H), 8.15 (s, 1H), 7.58 (m, 2H), 7.37 (dd, J=8.7,8.1 Hz, 2H), 5.60 (s, 2H), 3.92 (s, 4H), 3.82 (m, 2H), 2.89 (t, J=4.9Hz, 2H), 2.29 (s, 3H), 2.01 (m, 2H); MS: (ES) m/z calculated forC₂₁H₂₂ClFN₇O [M+H]; 442.2, found 442.1.

Example 70: Synthesis of2-[4-chloro-3-(1H-imidazol-2-yl)-5-methyl-pyrazol-1-yl]-1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]ethanone

A mixture of2-[4-chloro-3-(4,5-dihydro-1H-imidazol-2-yl)-5-methyl-pyrazol-1-yl]-1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]ethanone(45 mg, 0.1 mmol), Dess-Martin periodinane (80 mg, 0.2 mmol) in DMSO (1mL) was heated at 80° C. for 1 hour. After cooling to room temperature,the reaction mixture was diluted with CH₂Cl₂ (20 mL) and washed withwater (3×5 mL). The organic layer was dried (Na₂SO₄), filtered,concentrated in vacuo and purified by reverse phase HPLC (C18 column,acetonitrile-H₂O with 0.1% TFA as eluent) to give2-[4-chloro-3-(1H-imidazol-2-yl)-5-methyl-pyrazol-1-yl]-1-[1-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]ethanoneas white powder (32 mg, 73%). ¹H NMR (400 MHz, DMSO-d₆) δ 8.09 (s, 1H),7.60 (s, 2H), 7.52 (m, 2H), 7.30 (dd, J=8.8, 8.3 Hz, 2H), 5.49 (s, 2H),3.90 (m, 2H), 2.82 (t, J=5.1 Hz, 2H), 2.22 (s, 3H), 1.95 (m, 2H); MS:(ES) m/z calculated for C₂₁H₂₀ClFN₇O [M+H]⁺ 440.1, found 440.1.

Example 71: Synthesis of(2S)-1-[l-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-[2-methyl-4-(trifluoromethyl)imidazol-1-yl]propan-1-one

To a solution of(2S)-2-[2-methyl-4-(trifluoromethyl)imidazole-1-yl]propanoic acid (0.020g, 0.09 mmol) in CH₂Cl₂ (3 mL) was added oxalyl chloride (0.040 mL, 0.46mmol) and DMF (1 drop). After 20 min at room temperature, the mixturewas concentrated in vacuo and the residue was added to another flaskcontaining 1-(4-fluorophenyl)-4,5,6,7-tetrahydropyrazolo[4,3-b]pyridine(0.028 g, 0.13 mmol) and NEt₃ (0.060 mL, 0.43 mmol) in CH₂Cl₂ (3 mL).The resulting mixture was stirred at room temperature for 20 min,quenched with saturated aqueous NaHCO₃ solution (30 mL), and extractedwith ethyl acetate (50 mL). The organic layer was dried over anhydroussodium sulfate, concentrated in vacuo, and purified by flashchromatography (SiO₂, 0-8° % MeOH/EtOAc) to afford the titled compound(0.030 g, 44%, TFA salt) as a white solid. ¹H NMR (TFA salt) (400 MHz,CDCl₃) δ 9.70 (s, 1H), 8.43 (s, 1H), 7.78 (d, J=0.8 Hz, 1H), 7.42 (dd,J=9.2, 8.8 Hz, 2H), 7.20 (dd, J=8.8, 8.4 Hz, 2H), 5.61 (q, J=7.2 Hz,1H), 3.89 (m, 2H), 2.84 (dd, J=6.2, 6.2 Hz, 2H), 2.65 (s, 3H), 2.15 (m,2H), 1.84 (d, J=7.2 Hz, 3H); MS: (ES) m/z calculated for C₂₀H₉F₄N₅O[M+H]⁺ (free form) 422.1, found 422.1; The titled compounds wereanalyzed by chiral normal phase chromatography (RegisPack, 25 cm×4.6 mm,5 micron, cat#793104, 0.1% DEA/IPA, 0.7 mL/min). The (S)-enantiomer(major) had a retention time of 7.2 min and the (R)-enantiomer (minor)had a retention time of 6.2 min (isolated in 19:1 er).

Example 72: Synthesis of(2S)-1-[I-(4-chlorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-[2-methyl-4-(trifluoromethyl)imidazol-1-yl]propan-1-one

To a solution of(2S)-2-[2-methyl-4-(trifluoromethyl)imidazole-1-yl]propanoic acid (0.020g, 0.09 mmol) in CH₂Cl₂ (1.5 mL) was added oxalyl chloride (0.050 mL,0.58 mmol) and DMF (1 drop). After 20 min at room temperature, themixture was concentrated in vacuo and the residue was added to anotherflask containing1-(4-chlorophenyl)-4,5,6,7-tetrahydropyrazolo[4,3-h]pyridine (0.028 g,0.13 mmol) and NEt₃ (0.050 mL, 0.35 mmol) in CH₂Cl₂ (1 mL). Theresulting mixture was stirred at room temperature for 20 min, quenchedwith saturated aqueous NaHCO₃ solution (30 mL), and extracted with ethylacetate (50 mL). The organic layer was dried over anhydrous sodiumsulfate, concentrated in vacuo, and purified by flash chromatography(SiO₂, 0-6% MeOH/EtOAc) to afford the titled compound (0.022 g, 32%, TFAsalt) as a white solid. ¹H NMR (free form) (400 MHz, CDCl₃) δ 8.46 (s,1H), 7.44 (m, 4H), 7.42 (dd, J=9.2, 8.8 Hz, 2H), 7.37 (d, J=1.2 Hz, 1H),5.21 (q, J=7.2 Hz, 1H), 3.75 (m, 1H), 3.53 (m, 1H), 2.86 (dd, J=6.4, 6.4Hz, 2H), 2.46 (s, 3H), 1.72 (d, J=6.8 Hz, 3H); MS: (ES) m/z calculatedfor C₂₀H₁₉ClF₃N₅O [M+H]⁺ (free form) 438.1, found 438.1; The titledcompounds were analyzed by chiral normal phase chromatography(RegisPack, 25 cm×4.6 mm, 5 micron, cat#793104, 0.10% DEA/IPA, 0.7mL/min). The (S)-enantiomer (major) had a retention time of 8.4 min andthe (R)-enantiomer (minor) had a retention time of 6.4 min (isolated in18:1 er).

Example 73: Synthesis of1-[I-(4-fluorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-h]pyridin-4-yl]-2-[5-methyl-3-(trifluoromethyl)-1,24-triazol-1-yl]propan-1-one

a) A solution of ethyl 2-bromopropionate (3.16 g, 17.5 mmol),5-methyl-3-(trifluoromethyl)-1H-1,2,4-triazole (2.20 g, 14.6 mmol), andK₂CO₃ (4.00 g, 29.1 mmol) in THF/H₂O (2:1, 30 mL) was stirred at roomtemperature for 5 h. The mixture was diluted in EtOAc (50 mL). Theorganic layer was washed with water and brine, dried (MgSO₄), filtered,and concentrated in vacuo to give the desired product (4.0 g) as acolorless oil that was used in the next step without furtherpurification.

b) The crude material from step a in THF (40 mL) was treated with 2 NLiOH (15 mL, 30 mmol) at 45° C. for 1 h and concentrated. The residuewas diluted with water (20 mL), adjusted to pH 2 with 1 M H₂SO₄, andextracted with EtOAc (50 mL). The organic layer was dried (MgSO₄),filtered, and concentrated in vacuo to give the desired acid (2.56 g,10.2 mmol, 70% over two steps) as a colorless solid.

c) To a mixture of2-[5-methyl-3-(trifluoromethyl)-1,2,4-triazol-1-yl]propanoic acid (0.051g, 0.23 mmol) and1-(4-fluorophenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-b]pyri dine (0.050g, 0.23 mmol) in DMF (1 mL) were added Hunig's base (0.059 g, 0.46 mmol)and 2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uraniumhexafluorophosphate methanaminium (HATU) (0.096 g, 0.25 mmol). Themixture was stirred at room temperature for 1 hour and then partitionedbetween water (4 mL) and ethyl acetate (6 mL). The organic layer waswashed with brine, dried (Na₂SO₄), filtered, concentrated in vacuo, andpurified by reverse phase HPLC (C18 column, acetonitrile-H₂O with 0.1%TFA as eluent) to give the desired product as a white solid (0.029 g,0.069 mmol, 30%). ¹H NMR (400 MHz, CDCl3) δ 8.42 (s, 1H), 7.44 (ddd,J=10.4, 5.2, 2.8 Hz, 2H), 7.21-7.10 (m, 2H), 5.61 (q, J=7.1 Hz, 1H);3.77 (ddd, J=12.6, 7.5, 3.0 Hz, 1H), 3.52 (ddd, J=12.5, 8.4, 3.0 Hz,1H), 2.82 (td, J=6.4, 2.4 Hz, 2H), 2.52 (s, 3H), 2.10-1.80 (m, 5H); MS:(ES) mm/z calculated for C₁₉H₁₈ClF₃N₆ O[M+H]⁺ 423.1, found 422.9.

Example 74: Synthesis of1-[1-(4-chlorophenyl)-6,7-dihydro-5H-pyrazolo[4,3-b]pyridin-4-yl]-2-[5-methyl-3-(trifluoromethyl)-1,2,4-triazol-1-yl]propan-1-one

To a mixture of2-[5-methyl-3-(trifluoromethyl)-1,2,4-triazol-1-yl]propanoic acid (0.048g, 0.21 mmol) and1-(4-chlorophenyl)-4,5,6,7-tetrahydro-H-pyrazolo[4,3-b]pyridine (0.050g, 0.21 mmol) in DMF (1 mL) were added Hunig's base (0.055 g, 0.42 mmol)and 2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uraniumhexafluorophosphate methanaminium (HATU) (0.090 g, 0.23 mmol). Themixture was stirred at room temperature for 1 hour, and then partitionedbetween water (4 mL) and ethyl acetate (6 mL). The organic layer waswashed with brine, dried (Na₂SO₄), filtered, concentrated in vacuo, andpurified by reverse phase HPLC (C18 column, acetonitrile-H₂O with 0.1%TFA as eluent) to give the desired product as a white solid (0.028 g,0.063 mmol, 30%). ¹H NMR (400 MHz, CDCl3) δ 8.44 (s, 1H), 7.43 (s, 4H),5.60 (q, J=7.1 Hz, 1H), 3.77 (ddd, J=12.6, 7.5, 3.0 Hz, 1H), 3.52 (ddd,J=12.3, 8.3, 3.0 Hz, 1H), 2.84 (tt, J=6.9, 3.3 Hz, 2H), 2.52 (s, 3H),2.09-1.81 (m, 5H); MS: (ES) m/z calculated for C₁₉H₁₈ClF₃N₆ O[M+H]⁺439.1, found 438.9.

Example 75

This example illustrates the the evaluation of the biological activityassociated with compounds of interest (candidate compounds) of theinvention.

Materials and Methods

A. Cells

1. CCR1 Expressing Cells

a) THP-1 Cells

THP-1 cells were obtained from ATCC (TIB-202) and cultured as asuspension in RPMI-1640 medium supplemented with 2 mM L-glutamine, 1.5g/L sodium bicarbonate, 4.5 g/L glucose, 10 mM HEPES, 1 mM sodiumpyruvate, 0.05% 2-mercaptoethanol and 10% FBS. Cells were grown under 5%CO₂/95% air, 100% humidity at 37° C. and subcultured twice weekly at 1:5(cells were cultured at a density range of 2×10⁵ to 2×10⁶ cells/mL) andharvested at 1×10⁶ cells/mL. THP-1 cells express CCR1 and can be used inCCR1 binding and functional assays.

b) Isolated Human Monocytes

Monocytes were isolated from human buffy coats using the Miltenyi beadisolation system (Miltenyi, Auburn, Calif.). Briefly, following a Ficollgradient separation to isolate peripheral blood mononuclear cells, cellswere washed with PBS and the red blood cells lysed using standardprocedures. Remaining cells were labeled with anti-CD14 antibodiescoupled to magnetic beads (Miltenyi Biotech, Auburn, Calif.). Labeledcells were passed through AutoMACS (Miltenyi, Auburn, Calif.) andpositive fraction collected. Monocytes express CCR1 and can be used inCCR1 binding and functional assays.

B. Assays

1. Inhibition of CCR1 Ligand Binding

CCR1 expressing cells were centrifuged and resuspended in assay buffer(20 mM HEPES pH 7.1, 140 mM NaCl, 1 mM CaCl₂), 5 mM MgCl₂, and with 0.2%bovine serum albumin) to a concentration of 5×10⁶ cells/mL for THP-1cells and 5×10⁵ for monocytes. Binding assays were set up as follows.0.1 mL of cells (5×10⁵ THP-1 cells/well or 5×10⁴ monocytes) was added tothe assay plates containing the compounds, giving a final concentrationof ˜2-10 μM each compound for screening (or part of a dose response forcompound IC₅₀ determinations). Then 0.1 mL of ¹²⁵I labeled MIP-1α(obtained from Perkin Elmer Life Sciences, Boston, Mass.) or 0.1 mL of¹²⁵I labeled CCL15/leukotactin (obtained as a custom radiolabeling byPerkin Elmer Life Sciences, Boston, Mass.) diluted in assay buffer to afinal concentration of ˜50 μM, yielding ˜30,000 cpm per well, was added(using ¹²⁵I labeled MIP-1α with THP-1 cells and ¹²⁵I labeledCCL15/leukotactin with monocytes), the plates sealed and incubated forapproximately 3 hours at 4° C. on a shaker platform. Reactions wereaspirated onto GF/B glass filters pre-soaked in 0.3% polyethyleneimine(PEI) solution, on a vacuum cell harvester (Packard Instruments;Meriden, Conn.). Scintillation fluid (40 μl; Microscint 20, PackardInstruments) was added to each well, the plates were sealed andradioactivity measured in a Topcount scintillation counter (PackardInstruments). Control wells containing either diluent only (for totalcounts) or excess MIP-1α or MIP-1β (1 μg/mL, for non-specific binding)were used to calculate the percent of total inhibition for compound. Thecomputer program Prism from GraphPad, Inc. (San Diego, Ca) was used tocalculate IC₅₀ values. IC₅₀ values are those concentrations required toreduce the binding of labeled MIP-1α to the receptor by 50%. (Forfurther descriptions of ligand binding and other functional assays, seeDairaghi, et al., J. Biol. Chem. 274:21569-21574 (1999), Penfold, etal., Proc. Natl. Acad. Sci. USA. 96:9839-9844 (1999), and Dairaghi, etal., J. Biol. Chem. 272:28206-28209 (1997)).

2. Calcium Mobilization

To detect the release of intracellular stores of calcium, cells (THP-1or monocytes) were incubated with 3 μM of INDO-1 AM dye (MolecularProbes; Eugene, Oreg.) in cell media for 45 minutes at room temperatureand washed with phosphate buffered saline (PBS). After INDO-1AM loading,the cells were resuspended in flux buffer (Hank's balanced salt solution(HBSS) and 1% FBS). Calcium mobilization was measured using a PhotonTechnology International spectrophotometer (Photon TechnologyInternational; New Jersey) with excitation at 350 nm and dualsimultaneous recording of fluorescence emission at 400 nm and 490 nm.Relative intracellular calcium levels were expressed as the 400 nm/490nm emission ratio. Experiments were performed at 37° C. with constantmixing in cuvettes each containing 10⁶ cells in 2 mL of flux buffer. Thechemokine ligands may be used over a range from 1 to 100 nM. Theemission ratio was plotted over time (typically 2-3 minutes). Candidateligand blocking compounds (up to 10 μM) were added at 10 seconds,followed by chemokines at 60 seconds (i.e., MIP-1α; R&D Systems;Minneapolis, Minn.) and control chemokine (i.e., SDF-1α; R&D Systems;Minneapolis, Minn.) at 150 seconds.

3. Chemotaxis Assays

Chemotaxis assays were performed using 5 μm pore polycarbonate,polyvinylpyrrolidone-coated filters in 96-well chemotaxis chambers(Neuroprobe; Gaithersburg, Md.) using chemotaxis buffer (Hank's balancedsalt solution (HBSS) and 1% FBS). CCR1 chemokine ligands (i.e., MIP-1α,CCL15/Leukotactin; R&D Systems; Minneapolis, Minn.) are use to evaluatecompound mediated inhibition of CCR1 mediated migration. Otherchemokines (i.e., SDF-1α; R&D Systems; Minneapolis, Minn.) are used asspecificity controls. The lower chamber was loaded with 29 μl ofchemokine (i.e., 0.1 nM CCL15/Leukotactin) and varying amounts ofcompound; the top chamber contained 100,000 THP-1 or monocyte cells in20 μl. The chambers were incubated 1-2 hours at 37° C., and the numberof cells in the lower chamber quantified either by direct cell counts infive high powered fields per well or by the CyQuant assay (MolecularProbes), a fluorescent dye method that measures nucleic acid content,and by measuring with a Spectrafluor Plus (Tecan). The computer programPrism from GraphPad, Inc. (San Diego, Ca) was used to calculate IC₅₀values. IC₅₀ values are those compound concentrations required toinhibit the number of cells responding to a CCR1 agonist by 50%.

4. In Vivo Efficacy

a) Rabbit Model of Destructive Joint Inflammation

A rabbit LPS study was conducted essentially as described in Podolin, etal. J. Immunol. 169(11):6435-6444 (2002). Female New Zealand rabbits(approximately 2 kilograms) were treated intra-articularly in both kneeswith LPS (10 ng). The compound of interest, for example 1.016,(formulated in 1% methocel) or vehicle (1% methocel) was dosed orally ata 5 ml/kg dose volume at two times (2 hours before the intra-articularLPS injection and 4 hours after the intra-articular LPS injection).Sixteen hours after the LPS injection, knees were lavaged and cellscounts were performed. Beneficial effects of treatment were determinedby reduction in the number of inflammatory cells recruited to theinflamed synovial fluid of the knee joints. Treatment with the compoundof interest resulted in a significant reduction in recruitedinflammatory cells.

b) Evaluation of a Compound of Interest in a Rat Model of CollagenInduced Arthritis

A 17 day developing type II collagen arthritis study is conducted toevaluate the effects of a compound of interest on arthritis inducedclinical ankle swelling. Rat collagen arthritis is an experimental modelof polyarthritis that has been widely used for preclinical testing ofnumerous anti-arthritic agents (see Trentham, et al., J. Exp. Med.146(3):857-868 (1977), Bendele, et al., Toxicologic Pathol. 27:134-142(1999), Bendele, et al., Arthritis Rheum. 42:498-506 (1999)). Thehallmarks of this model are reliable onset and progression of robust,easily measurable polyarticular inflammation, marked cartilagedestruction in association with pannus formation and mild to moderatebone resorption and periosteal bone proliferation.

Female Lewis rats (approximately 0.2 kilograms) are anesthetized withisoflurane and injected with Freund's Incomplete Adjuvant containing 2mg/mL bovine type II collagen at the base of the tail and two sites onthe back on days 0 and 6 of this 17 day study. A compound of interest isdosed daily in a sub-cutaneous manner from day 0 till day 17 at aefficacious dose. Caliper measurements of the ankle joint diameter aretaken, and reduced joint swelling is taken as a measure of efficacy.

Murine Model of Dermatological Disease

Compounds of the invention can be assessed in the murine model of dermaldelayed type hypersensitivity induced by oxazolone. Briefly, 8-10 weekold BALB/c mice are sensitized topically with a 1% solution of oxazolonedissolved in ethanol on their shaved abdomens on day 0. On day 6 postsensitization mice are dosed orally with either vehicle or increasingdoses of a compound of the invention immediately prior to and 4 hoursfollowing a topical challenge with a 0.5% solution of oxazolone inethanol on the right ear. The following day (day 7), ear thicknesses aremeasured using caliper measurements. Animals treated with compound havesignificantly reduced ear swelling compared to vehicle treated controlsindicating a compound mediated decrease in oxazolone induced dermalhypersensitivity.

Murine Asthma Model

Compounds of the invention can be assessed in the murine model ofallergic asthma. Asthma is induced in 8-10 week old BALB/c mice bysensitizing mice with OVA in Alum adjuvant on days 0 and 10. On day 20mice are challenged with OVA in PBS intranasally to elicit airwayinflammation. Groups of mice are either treated with vehicle, orincreasing doses of a compound of the invention starting on day 20 andlasting until day 23. Animals are analyzed at day 23 after theintranasal OVA challenge for cellular infiltrates in bronchoalveolarlavage (BAL). A significant reduction in BAL leukocyte numbers relativeto vehicle treated mice indicates the compound is effective in thismodel.

Murine Model of Cancer

This example describes a procedure to evaluate efficacy of CCR1antagonists for treatment of malignancy. Normal mouse strains can betransplanted with a variety of well-characterized mouse tumor lines,including a mouse thymoma EL4 which has been transfected with OVA toallow easy evaluation of tumor specific antigen responses followingvaccination with OVA. Three series of mouse groups from any of thesetumor models are tested for CCR1 antagonist efficacy as follows: Oneseries of mice additionally receives PBS and Tween 0.5% i.p. soon aftertumor transplant, and thereafter at varying dosing schedules. A secondseries consists of groups of mice receiving different doses of the CCR1antagonist given either intra-peritoneally, intra-venously,sub-cutaneously, intramuscularly, orally, or via any other mode ofadministration soon after tumor transplant, and thereafter at varyingdosing schedules. A third series of mice, serving as positive control,consists of groups treated with either anti-IL4 antibodies, anti-IFNgantibodies, IL4, or TNF, given i.p. soon after tumor transplant, andthereafter at varying dosing schedules. Efficacy is monitored via tumorgrowth versus regression. In the case of the OVA-transfected EL4 thymomamodel, cytolytic OVA-specific responses can be measured by stimulatingdraining lymph node cells with OVA in vitro, and measuringantigen-specific cytotoxicity at 72 hours.

Murine Model of Inflammatory Bowel Diseases

The MDR1a-knockout mice, which lack the P-glycoprotein gene,spontaneously develop colitis under specific pathogen-free condition.The pathology in these animals has been characterized as Th1-type Tcell-mediated inflammation similar to ulcerative colitis in humans.Disease normally begins to develop at around 8-10 weeks after birth.However the ages at which disease emerges and the ultimate penetrancelevel often vary considerably among different animal facilities. In astudy using the MDR1a-knockout mice, a CCR1 antagonist can be evaluatedprophylacticly or therapeutically depending on time of administration.Female mice (n=34) are dosed with a compound of interest as appropriateto the compound eg daily in a sub-cutaneous manner at a efficaciousdose. The study is evaluated for IBD associated growth retardation andscoring of anal discharge and irritation. A compound which reduces analdischarge and irritation or inhibits IBD associated growth retardationindicates efficacy of compound in this indication.

Murine Model of Solid Tumors

The mouse RENCA tumor model accurately mimics the progression of humanadult renal cell carcinoma specifically with reference to spontaneousmetastasis to lungs and serves as a model for solid tumors. Balb/c 6-8week old female mice are inoculated with approximately 5e5 RENCA cells(mouse renal adenocarcinoma; ATCC cat# CRL-2947) under the kidneycapsule and kidney tumor growth is observed over 22 days, with lungmetastasis observed as early as day 15. Animals are dosed with eithervehicle or a compound of the invention eg daily subcutaneously, from thetime of tumor implantation to monitor effects on primary growth, or at alater time (eg day 7) to monitor the compound effect on metastasis.Primary tumor areas are measured twice a week using mechanical calipers.Tumor volumes are calculated by the formula v=pab2/6, where a is thelongest diameter and b is the next longest diameter perpendicular to a.A reduction in tumor volume or incidence of metastasis indicatesefficacy of compound in this indication.

Murine Model of Radiation-Induced Pulmonary Disease (RIPD)

Various models can be used to assess the efficacy of a compound of theinvention in recovery from RIPD. For instance Tokuda et al describe amodel of bleomycin induced lung fibrosis (Tokuda et al, J Immunol.164(5):2745-51 (2000)). Alternatively Yang et al describe a model ofdirect radiation induced disease (Yang et al, Am J Respir Cell Mol Biol.45(1):127-35 (2011)). Briefly, anesthetized compound treated oruntreated mice aged 10-12 weeks are immobilized, all but the thoraciccavity lead shielded and animals irradiated with a single dose of 14.5Gy from a cesium source at 1.65 Gy/min. At this dose, survival issufficient to permit adequate numbers of animals for long-term analyses.Efficacy of compound can be assessed by many methods know in the art,including lung mechanical assessment, lung hydroxyproline levels andothers.

In the table below, structures and activity are provided forrepresentative compounds described herein. Activity is provided asfollows for either the chemotaxis assay or binding assay as describedabove: +, IC₅₀>12.5 uM; ++, 2500 nM<IC₅₀<12.5 uM; +++, 1000 nM<IC₅₀<2500nM; and ++++, IC₅₀<1000 nM.

TABLE 1 Specific Examples CTX IC50 Example (nM)

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1.-27. (canceled)
 28. A method of assaying a compound for CCR1antagonistic activity, said method comprising (a) contacting thecompound with cells expressing CCR1 and a radioactive CCR1 ligand toform a reaction mixture; (b) aspirating the reaction mixture onto a GF/Bglass filter pre-soaked in a polyethyleneimine solution; (c) measuringthe amount radioactivity remaining on the GF/B glass filter, whereinsaid method comprises performing steps (a)-(c) with a positive controlsample having a formula represented by the structure

wherein the subscript n is an integer of from 0 to 3; each R^(1a) andR^(b) is a member independently selected from the group consisting of H,C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₃₋₆ cycloalkyl, —COR^(a), —CO₂R^(a),—CONR^(a)R^(b), —NR^(a)R^(b), —NR^(a)COR^(b), —OR^(a), —X¹COR^(a),—X¹CO₂R^(a), —X¹CONR^(a)R^(b), —X¹NR^(a)COR^(b), —X¹NR^(a)R^(b), and—X¹OR^(a), wherein X¹ is a member selected from the group consisting ofC₁₋₄ alkylene, and each R^(a) and R^(b) is independently selected fromthe group consisting of hydrogen, C₁₋₈ alkyl, C₁₋₈ haloalkyl, and C₃₋₆cycloalkyl, and optionally two R^(1a) groups on adjacent carbon atomsare joined to form a 5-, 6- or 7-membered carbocyclic or heterocyclicring; each of R^(2a) and R^(2b) is a member independently selected fromthe group consisting of H, hydroxyl, C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₁₋₈alkoxy, C₁₋₄ alkoxy-C₁₋₄ alkyl, C₁₋₈ hydroxyalkyl, C₁₋₄ alkoxy-C₁₋₄alkoxy, C₃₋₆ cycloalkyl, C₃₋₆ cycloalkyl-C₁₋₄ alkyl, 3- to 7-memberedheterocycloalkyl, 3- to 7-membered heterocycloalkyl-C₁₋₄ alkyl,—X¹CO₂R^(a), —X¹CONR^(a)R^(b), —X¹NR^(a)COR^(b), —X¹NR^(a)R^(b), whereinX¹, R^(a) and R^(b) are defined above; Ar¹ is a member selected from thegroup consisting of phenyl, naphtyl and pyridyl, each of which issubstituted with from one to five substituents, R³, R^(3a), R^(3b), R⁴and R^(4a) which are independently selected from the group consisting ofH, halogen, —OR^(c), —OC(O)R^(c), —NR^(c)R^(d), —SR^(c), —R^(c), —CN,—NO₂, —CO₂R^(c), —CONR^(c)R^(d), —C(O)R^(c), —OC(O)NR^(c)R^(d),—NR^(d)C(O)R^(c), —NR^(d)C(O)₂R, —NR^(c)—C(O)NR^(c)R^(d), —NH—C(NH₂)═NH,—NR^(e)C(NH₂)═NH, —NH—C(NH₂)═NR^(e), —NH—C(NHR^(e))═NH, —S(O)R,—S(O)₂R^(e), —NR^(c)S(O)₂R^(e), —S(O)₂NR^(c)R^(d), —N₃, —X²OR^(c),—O—X²OR^(c), —X²OC(O)R^(c), —X²NR^(c)R^(d), —O—X²NR^(c)R^(d), —X²SR^(c),—X²CN, —X²NO₂, —X²CO₂R^(c), —O—X²CO₂R^(c), —X²CONR^(c)R^(d),—O—X²CONR^(c)R^(d), —X²C(O)R^(c), —X²OC(O)NR^(c)R^(d),—X²NR^(d)C(O)R^(c), —X²NR^(d)C(O)₂R^(e), —X²NR^(c)C(O)NR^(c)R^(d),—X²NH—C(NH₂)═NH, —X²NR^(e)C(NH₂)═NH, —X²NH—C(NH₂)═NR^(e),—X²NH—C(NHR^(e))═NH, —X²S(O)R, —X²S(O)₂R, —X²NR^(c)S(O)₂R^(e),—X²S(O)₂NR^(c)R^(d), —X²N₃, —NR^(d)—X²OR, —NR^(d)—X²NR^(c)R^(d),—NR^(d)—X²CO₂R^(c), and —NR^(d)—X²CONR^(c)R^(d), wherein each X² is amember independently selected from the group consisting of C₁₋₄alkylene, and each R^(c) and R^(d) is independently selected fromhydrogen, C₁₋₈ alkyl, C₁₋₈ hydroxyalkyl, C₁₋₈ haloalkyl and C₃₋₆cycloalkyl, or optionally R and R^(d) when attached to the same nitrogenatom can be combined with the nitrogen atom to form a five orsix-membered ring having from 0 to 2 additional heteroatoms as ringmembers; and each R is independently selected from the group consistingof C₁₋₈ is alkyl, C₁₋₈ hydroxyalkyl, C₁₋₈ haloalkyl and C₃₋₆ cycloalkyl;Ar² is a member selected from the group consisting of a six- orten-membered monocyclic or fused bicyclic aryl ring, and a five- toten-membered monocyclic or fused bicyclic heteroaryl ring; each of whichis substituted with from one to five substituents, R⁵, R⁶, R⁷, R⁸ andR⁹, independently selected from the group consisting of H, halogen,—OR^(f), —OC(O)R^(f), —NR^(f)R^(g), —SR, —R^(h), —CN, —NO₂, —CO₂R^(f),—CONR^(f)R^(g), —C(O)R^(f), —OC(O)NR^(f)R^(g), —NR^(g)C(O)R^(f),—NR^(g)C(O)₂R^(h), —NR^(f)—C(O)NR^(f)R^(g), —NH—C(NH₂)═NH,—NR^(h)C(NH₂)═NH, —NH—C(NH₂)═NR^(h), —NH—C(NHR^(h))═NH, —S(O)R^(h),—S(O)₂R^(h), —NR^(f)S(O)₂R^(h), —S(O)₂NR^(f)R^(g),—NR^(f)S(O)₂NR^(f)R^(g), —N₃, —X³OR, —X³OC(O)R^(f), —X³NR^(f)R^(g),—X³SR^(f), —X³CN, —X³NO₂, —X³CO₂R, —X³CONR^(f)R^(g), —X³C(O)R^(f),—X³OC(O)NR^(f)R^(g), —X³NR^(g)C(O)R^(f), —X³NR^(g)C(O)₂R^(h),—X³NR^(f)—C(O)NR^(f)R^(g), —X³NH—C(NH₂)═NH, —X³NR^(h)C(NH₂)═NH,—X³NH—C(NH₂)═NR^(h), —X³NH—C(NHR^(h))═NH, —X³S(O)R^(h), —X³S(O)₂R^(h),—X³NR^(f)S(O)₂R^(h), —X³S(O)₂NR^(f)R^(g), —Y, —X³Y, —S(O)₂Y, —C(O)Y,—X³N₃, —O—X³OR^(f), —O—X³NR^(f)R^(g), —O—X³CO₂R, —O—X³CONR^(f)R^(g),—NR^(g)—X³OR^(f), —NR^(g)—X³NR^(f)R^(g), —NR^(g)—X³CO₂R^(f), and—NR⁹—X³CONR^(f)R^(g), wherein Y is a five or six-membered aryl,heteroaryl or heterocyclic ring, optionally substituted with from one tothree substitutents selected from the group consisting of halogen,—OR^(f), —OC(O)R, —NR^(f)R^(g), —R^(h), —SR^(f), —CN, —NO₂, —CO₂R^(f),—CONR^(f)R^(g), —C(O)R^(f), —NR^(g)C(O)R^(f), —NR^(g)C(O)₂R^(h),—S(O)R^(h), —S(O)₂R^(h), —NR^(f)S(O)₂R^(h), —S(O)₂NR^(f)R^(g),—X³OR^(f), —X³SR^(f), —X³CN, —X³NO₂, —X³CO₂R^(f), —X³CONR^(f)R^(g),—X³C(O)R^(f), —X³OC(O)NR^(f)R^(g), —X³NR^(g)C(O)R^(f),—X³NR^(g)C(O)₂R^(h), —X³NR^(f)—C(O)NR^(f)R^(g), —X³OC(O)R^(f),—X³S(O)R^(h), —X³S(O)₂R^(h), —X³NR^(f)R^(g), —X³NR^(f)S(O)₂R^(h),—X³S(O)₂NR^(f)R^(g), —O—X³OR^(f), —O—X³NR^(f)R^(g), —O—X³CO₂R,—O—X³CONR^(f)R^(g), —NR^(g)—X³OR^(f), —NR^(g)—X³NR^(f)R^(g),—NR^(g)—X³CO₂R^(f), and —NR^(g)—X³CONR^(f)R^(g) and wherein each X³ isindependently selected from the group consisting of C₁₋₄ alkylene, andeach R^(f) and R^(g) is independently selected from hydrogen, C₁₋₈alkyl, C₁₋₈ hydroxyalkyl, C₁₋₈ haloalkyl and C₃₋₆ cycloalkyl, or whenattached to the same nitrogen atom can be combined with the nitrogenatom to form a five or six-membered ring having from 0 to 2 additionalheteroatoms as ring members, and each R^(h) is independently selectedfrom the group consisting of C₁₋₈ alkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl,C₁₋₈ hydroxyalkyl, C₁₋₈ haloalkyl and C₃₋₆ cycloalkyl; or when two ofR⁵, R⁶, R⁷, R⁸ and R⁹, are attached to adjacent ring vertices of Ar²,are optionally combined to form a five or six membered ring having zero,one or two heteroatoms selected from O and N as ring members; or apharmaceutically acceptable salt, or rotamer thereof.
 29. The method ofclaim 28, wherein said cells expressing CCR1 are THP-1 cells or isolatedhuman monocytes.
 30. The method of claim 28, wherein said radioactiveCCR1 ligand is MIP-1α, MPIF-1, Leukotactin, or a combination thereof.31. The method of claim 28, wherein said reaction mixture comprises anassay buffer comprising 25 mM Hepes, 500 mM NaCl, 1 mM CaCl₂, 5 mMMgCl₂, pH 7.1.
 32. The method of claim 28, wherein said measuringcomprises adding scintillation fluid to the aspirated GF/C glass filter.33. The method of claim 28, wherein Ar² is selected from the groupconsisting of phenyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl,oxazolyl, isoxazolyl, oxadiazolyl, oxathiadiazolyl, pyrrolyl, thiazolyl,isothiazolyl, benzimidazolyl, benzoxazolyl, benzopyrazolyl,benzotriazolyl, pyrazolo[3,4-b]pyridine, pyrazolo[3,4-d]pyrimidine,imidazo[4,5-b]pyridine, imidazo[1,5-α]pyridine, andpyrrolo[2,3-b]pyridine, each of which is optionally substituted with R⁵,R⁶ and R⁷.
 34. The method of claim 28, wherein Ar² is selected from thegroup consisting of pyrazolyl, imidazolyl and triazolyl, each of whichis substituted with R⁵, R⁶ and R⁷.
 35. The method of claim 28, Ar¹ isselected from the group consisting of phenyl, naphthyl and pyridyl, eachof which is substituted with from one to five substituents, R³, R^(3a),R^(3b), R⁴ and R^(4a); and Ar² is selected from the group consisting ofpyrazolyl, imidazolyl and triazolyl, each of which is substituted withR⁵, R⁶ and R⁷.
 36. The method of claim 28, wherein Ar¹ is phenyl, whichis substituted with from one to five substituents, R³, R^(3a), R^(3b),R⁴ and R^(4a), and Ar² is selected from the group consisting ofpyrazolyl, imidazolyl, benzimidazolyl, benzopyrazolyl,pyrazolo[3,4-b]pyridine, pyrazolo[3,4-d]pyrimidine,imidazo[4,5-b]pyridine, imidazo[1,5-α]pyridine, andpyrrolo[2,3-b]pyridine, each of which is optionally substituted with R⁵,R⁶ and R⁷.
 37. The method of claim 28, wherein the positive controlsample has a formula represented by the structure:

wherein R³ and R⁴ are independently selected from the group consistingof H, halogen, —R^(e), —CN, and —SO₂R.
 38. The method of claim 37,wherein Ar² is a heteroaryl group.
 39. The method of claim 37, whereinAr² is a heteroaryl group, optionally substituted and attached to theremainder of the molecule through a nitrogen atom ring vertex.
 40. Themethod of claim 39, wherein said Ar² has the formula:

wherein R⁵, R⁶, and R⁷ are independently selected from the groupconsisting of H, halogen, —R^(h), —CN, —SO₂R^(h), —CO₂R^(f),—CONR^(f)R^(g), and Y.
 41. The method of claim 40, the positive controlsample has a formula represented by the structure:

wherein R⁴ is selected from the group consisting of F and C₁.
 42. Themethod of claim 41, wherein Y is selected from the group consisting ofpyridyl, pyrimidinyl, imidazolyl, oxazolyl, oxadiazolyl, triazolyl,thiazolyl, imidazolinyl and pyrazolyl.
 43. The method of claim 41, thepositive control sample has a formula represented by the structure:

wherein R³ is selected from the group consisting of H, halogen, C₁₋₈alkyl, C₁₋₈ haloalkyl and C₁₋₈ alkoxy; R^(1a) and R² are independentlyselected from the group consisting of H, C₁₋₈ alkyl, C₁₋₈ haloalkyl,C₁₋₈ alkoxy and C₁₋₈ hydroxyalkyl.
 44. The method of claim 28, thepositive control sample has a formula represented by the structure:

wherein R^(1a) and R^(2a) are independently selected from the groupconsisting of H, C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₁₋₈ alkoxy and C₁₋₈hydroxyalkyl; and R⁵, R⁶, and R⁷ are independently selected from thegroup consisting of H, halogen, —R^(h), —CN, —SO₂R^(h),—CO₂R^(f)—CONR^(f)R^(g), and Y.
 45. The method of claim 28, the positivecontrol sample has a formula represented by the structure:

Where R^(1a) and R^(2a) are independently selected from the groupconsisting of H, C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₁₋₈ alkoxy and C₁₋₈hydroxyalkyl; and R⁵, R⁶, and R⁷ are independently selected from thegroup consisting of H, halogen, —R^(h), —CN, —SO₂R^(h),—CO₂R^(f)—CONR^(f)R^(g), and Y.
 46. The method of claim 28, the positivecontrol sample has a formula represented by the structure:

wherein R^(1a) and R^(2a) are each independently selected from the groupconsisting of H, C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₁₋₈ alkoxy, and C₁₋₈hydroxyalkyl; and R⁵ and R⁷ are each independently selected from thegroup consisting of H, halogen, —R^(h), —CN, —SO₂R^(h), —CO₂R^(f),—CONR^(f)R^(g), and Y.
 47. The method of claim 28, the positive controlsample has a formula selected from the group consisting of